N-(arylsulphonyl)amino acid derivatives, process for their preparation and pharmaceutical compositions containing them

ABSTRACT

The invention relates compounds of formula in which R1 to R9, R16 and R17 are as defined in claim 1. These compounds are pharmacologically active.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.09/101,214, filed Jul. 2, 1998 now U.S. Pat. No. 6,015,812, which is theU.S. national phase of PCT application PCT/FR97/00026, filed Jan. 7,1997, which claimed priority of French application 9600269, filed Jan.11, 1996. The entire disclosure of each of these documents isincorporated herein by reference.

The present invention relates to novel N-(arylsulphonyl)amino acidderivatives, to their preparation and to pharmaceutical compositionscontaining them.

These compounds have affinity for bradykinin (BK) receptors. Bradykininis a nonapeptide belonging, like the decapeptide kallidin, to the classof kinins and which shows physiological activity in the cardiovascularfield and as a mediator in inflammation and pain. Several bradykininreceptors are distinguished: the B₁ and B₂ receptors (D. Regoli et, al.,Pharmacol. Rev., 1980, 32, 1-46). More precisely, the B₂ receptors arethe bradykinin and kallidin receptors: they are predominant and arenormally found in most tissues; the B, receptors are the receptorsspecific for [des-Arg⁹ ] bradykinin and for [des-Arg¹⁰ ] kallidin: theyare induced during inflammatory processes.

Bradykinin receptors have been cloned for different species, inparticular for the human species: B₁ receptor : J. G. Menke et al., J.Biol. Chem., 1994, 269 (34) 21583-21586; B₂ receptor : J. F. Hess,Biochem. Biophys. Res. Commun., 1992,,184, 260-268.

The reviews: Drug News and Perspectives, 1994, 7 (10), 603-611 and Exp.Opin. Ther. Patents, 1995, 5 (4), 331-340, give an account onbradykinin-receptor antagonists. Many antagonists described have peptidestructures. As bradykinin-receptor antagonists, mention may be made inparticular of HOE-140 (F. J. Hock, Brit. J. Pharmacol. 1991, 102,769-773) for the B₂ receptor and [des-Arg⁹, Leu⁸ ] bradykinin for the B₁receptor (M. N. Perkins et al., Pain, 1993, 53, 191-197). Recently, a B₂receptor antagonist of non-peptide structure, SR 173657, has beendescribed in Archiv Pharmacol., 1996, Suppl. 1, 354 (4), R6.

According to the present invention, a novel family of compounds havingaffinity for the bradykinin receptors has now been found; thesecompounds are N-(arylsulphonyl)amino acid derivatives.

Among the N-(arylsulphonyl)amino acid derivatives, some are known andhave various pharmacological activities. Thus, compounds withanti-thrombotic activity are described in the European, German andinternational patents or patent applications EP 558,961, EP 236,163, EP236,164, DD 155,954, DE 4,115,468 and WO 92/16549. In this field ofactivity, NAPAP, derived from N-(naphthalenesulphonyl)-glycine offormula: ##STR2## is described in Pharmazie, 1987, 42 (5), 346. ##STR3##

Furthermore, N-tosyl-β-alanine derivatives of formula:

in which a and b, together with the nitrogen atom to which they areattached, constitute a ring such as piperidine, pyrrolidine ormorpholine, are described in Pharmazie, 1984, 39 (5), 315-317.

Similarly, N-(arylsulphonyl)proline derivatives have been cited asthrombin inhibitors in Pharmazie, 1986, 41 (4), 233-235 and Pharmazie,1987, 42 (2), 114-116.

Moreover, patent application EP 614,911 describes compounds of formula:##STR4## in which, in particular: Ar_(I) is a naphthyl, a phenyl, aquinolyl or an isoquinolyl, which are optionally substituted;

Ar_(II) is a phenyl or a thienyl, which are optionally substituted;

R_(I), R_(II) and R_(III) are, independently of each other, H or (C₁-C₄)alkyl;

or R_(I) is nothing and N is linked to Ar_(II) and optionally R_(II) andR'_(II) form a double bond;

or R_(I) or R_(II) is linked to Ar_(II) and is a (C₁ -C₃) alkylene;

R_(III) and R_(IV), which may be identical or different, are H, (C₁-C₄)alkyl or form, together with the nitrogen atom to which they areattached, a (C₅ -C₇) heterocycle;

Z₁ is a (C₁ -C₁₂)alkylene;

Q₁ is methyl, amino, (C₁ -C₄)alkoxycarbonylamino, (C₁ -C₄) alkylamino,di(C₁ -C₄)alkylamino, pyrrolidinyl, piperidino, morpholino, piperazinyl,(C₁ -C₄)alkyl-4-piperazinyl, amidino, (C₁ -C₄)alkylamidino, guanidino,(C₁ -C₄)alkylguanidino, pyridyl, imidazolyl, pyrimidinyl, indolyl,hydroxyl, (C₁ -C₄)alkoxy, (C₂ -C₈)alkoxycarbonyl, amino(C₁-C₄)alkyl-N-(C₁ -C₄)alkylamino, carbamoyl or phenyl, which is optionallysubstituted,

Q₂ is H or (C₁ -C₄)alkyl;

Q₃ is H or (C₁ -C₄)alkyl or Q₁ and Q₃ are linked to form a heterocycleand together are (C₂ -C₃)alkylene when Z₁ is nothing, in the form ofpure enantiomers or mixtures thereof in any proportion;

as well as the salts thereof with acids.

These compounds have affinity for the biological receptors of theneuropeptide Y.

According to the present invention, novel compounds have now been foundwhich have, unexpectedly, affinity for the bradykinin receptors.

The subject of the present invention is the compounds of formula:##STR5## in which: R₁ is a phenyl, a naphthyl, a tetrahydronaphthyl, aquinolyl or an isoquinolyl, the said rings being unsubstituted orsubstituted one or more times with R₁₀ ;

R₂ is a phenyl which is unsubstituted or substituted one or more timeswith R₁₁, a phenyl(C₁ -C₄)alkyl which is unsubstituted or substitutedone or more times on the phenyl with R₁₁, a naphthyl which isunsubstituted or substituted one or more times with R₁₁, a cyclohexylwhich is unsubstituted or substituted one or more times with R₁₁ ;

or R₂ and R₉ are linked together and constitute a (C₃ -C₅)alkylene whichis unsubstituted or substituted with R₁₂ or a (C₂ -C₄)alkylene which isinterrupted with an oxygen atom or a sulphur atom and is unsubstitutedor substituted with R₁₂ ;

or R₂ and R₉, together with the carbon atom and the nitrogen atom towhich they are attached, constitute tetrahydroisoquinoline which isunsubstituted or substituted one or more times with a halogen, ahydroxyl, a (C₁ -C₄)alkyl, a (C₁ -C₄)alkoxy or a benzyloxy;

R₃ is hydrogen or a hydroxyl;

R₄ and R₅ are each independently hydrogen or a (C₁ -C₄) alkyl;

or R₄ and R₅, together with the nitrogen atom to which they areattached, constitute a heterocyclic radical chosen from 1-pyrrolidinyl,1-piperidyl, perhydro-1-azepinyl, 4-morpholinyl, 4-oxo-1-piperidyl,dihydro-1-pyrrolyl or dihydro-2-imidazolyl, the said heterocyclicradicals being unsubstituted or substituted one or more times with R₁₃ ;

R₆ is hydrogen and R₆ can also be R₈ when R₇ is hydrogen;

R₇ is hydrogen or a (C₁ -C₄)alkyl;

R₈ is hydrogen; a benzyl which is unsubstituted or substituted on thephenyl one or more times with R₁₃ ; or a group ZR₁₄ ;

or R₇ and R₈, together with the nitrogen atom to which they areattached, constitute a heterocyclic radical chosen from 1-pyrrolidinyl,1-piperidyl, 1-perhydro-1-azepinyl, 4-morpholinyl,tetrahydro-2-pyrimidinyl, 1-piperazinyl or 1-piperazinyl substituted inposition 4 with a (C₁ -C₄)alkyl or a benzyl;

or, when R₇ is hydrogen, R₆ and Re are linked together to form a (C₂-C₄)alkylene which is unsubstituted or substituted one or more timeswith a (C₁ -C₄)alkyl;

R₉ is hydrogen, a (C₁ -C₄)alkyl or a phenyl(C₁ -C₄)alkyl which isunsubstituted or substituted on the phenyl one or more times with R₁₁ ;

R₁₀ is a halogen, a (C₁ -C₄)alkyl, a (C₁ -C₄)alkoxy, a hydroxyl, anamino, a (C₁ -C₄)alkylamino or a di(C₁ -C₄) alkylamino;

R₁₁ is a halogen, a (C₁ -C₄)alkyl, a trifluoromethyl, a phenyl, ahydroxyl, a (C₁ -C₄)alkoxy or a benzyloxy;

or R₁₁ is in the ortho position to the phenyl representing R₂ and formswith R₃ a methylene group or an ethylene group;

or R₁₁ is in the ortho position to the phenyl representing R₂ and formswith R₉ a methylene group or an ethylene group;

R₁₂ is a halogen, a (C₁ -C₄)alkyl, a hydroxyl, a (C₁ -C₄)alkoxy, abenzyloxy, an oxo, a phenyl, an acetyloxy or a trifluoroacetyloxy;

R₁₃ is a (C₁ -C₄)alkyl, a halogen or a hydroxyl;

R₁₄ is a methyl, an amino, a (C₁ -C₄)alkylamino, a di (C₁-C₄)alkylamino, a tri (C₁ -C₄)alkylammonium, an amidino, a (C₁-C₄)alkylamidino, a guanidino, a (C₁ -C₄)alkylguanidino, a hydroxyl, a(C₁ -C₄)alkoxy, a (C₁ -C₄)alkoxycarbonyl, a group--AlkN(R₁₅)Alk'N(R'₁₅)₂, or a heterocyclic radical chosen from1-pyrrolidinyl, 1-piperidyl, perhydro-1-azepinyl, pyridyl, imidazolyl,dihydroimidazolyl, imidazolidinyl, pyrimidinyl and indolyl;

R₁₅ and R'₁₅ are, independently of each other, hydrogen or a (C₁-C₄)alkyl;

R₁₆ is hydrogen or a methyl, or R₁₆ forms with R₉ a methylene group;

R₁₇ is hydrogen or a methyl;

Alk and Alk' are, independently of each other, a (C₁ -C₄)alkylene;

Z is a (C₂ -C₁₂)alkylene or a (C₁ -C₆)alkylene which is interrupted orsubstituted with a (C₅ -C₇)cycloalkyl or with a phenyl;

C* is an asymmetric carbon atom;

as well as the salts thereof with inorganic or organic acids.

The salts are generally prepared with pharmaceutically acceptable acids,but the salts of other acids which are useful for the purification orisolation of the compounds of formula (I) also form part of theinvention. The pharmaceutically acceptable salts of the compounds offormula (I) are, for example, the hydrochloride, the hydrobromide, thesulphate, the methanesulphonate, the benzenesulphonate, thenaphthalenesulphonate, the maleate, the fumarate, the citrate, theacetate, the gluconate, the dobesilate or the sultosilate.

The compounds of formula (I) comprise 2 (or possibly more) asymmetriccarbon atoms and the 4 (or possibly more) pure enantiomers, as well asthe mixture thereof in any proportion, are subjects of the invention.

The term halogen is understood to refer to chlorine, fluorine, bromineor iodine, chlorine and fluorine being preferred.

The terms, alkyl, alkylene and alkoxy are understood to refer,respectively, to a linear or branched alkyl radical, alkylene radical oralkoxy radical.

Those compounds of formula (I) are preferred in which:

R₁ is a phenyl, a naphthyl, a tetrahydronaphthyl, a quinolyl or anisoquinolyl, the said rings being unsubstituted or substituted one ormore times with R₁₀ ;

R₂ is a phenyl which is unsubstituted or substituted one or more timeswith R₁₁, a phenyl(C₁ -C₄)alkyl which is unsubstituted or substitutedone or more times on the phenyl with R₁₁, or a naphthyl which isunsubstituted or substituted one or more times with R₁₁ ;

or R₂ and R₉ are linked together and constitute a (C₃ -C₅)alkylene whichis unsubstituted or substituted with R₁₂ or a (C₂ -C₄)alkylene which isinterrupted with an oxygen atom or a sulphur atom and is unsubstitutedor substituted with R₁₂ ;

or R₂ and R₉ together with the carbon atom and the nitrogen atom towhich they are attached, constitute tetrahydroisoquinoline which isunsubstituted or substituted one or more times with a halogen, ahydroxyl, a (C₁ -C₄)alkyl, a (C₁ -C₄)alkoxy or a benzyloxy;

R₃ is hydrogen or a hydroxyl;

R₄ and R₅ are each independently hydrogen or a (C₁ -C₄) alkyl;

or R₄ and R₅, together with the nitrogen atom to which they areattached, constitute a heterocyclic radical chosen from 1-pyrrolidinyl,1-piperidyl, perhydro-1-azepinyl, 4-morpholinyl or 4-oxo-1-piperidyl,the said heterocyclic radicals being unsubstituted or substituted withR₁₃ ;

R₆ is hydrogen, R₆ can also be R₈ when R₇ is hydrogen;

R₇ is hydrogen or a (C₁ -C₄)alkyl;

R₈ is hydrogen; a benzyl which is unsubstituted or substituted on thephenyl one or more times with R₁₃ ; or a group ZR₁₄ ;

or R₇ and R₈, together with the nitrogen atom to which they areattached, constitute a heterocyclic radical chosen from 1-pyrrolidinyl,1-piperidyl, perhydro-1-azepinyl, 4-morpholinyl, 1-piperazinyl or1-piperazinyl substituted in position 4 with a (C₁ -C₄)alkyl or abenzyl;

or, when R₇ is hydrogen, R₆ and R₈ are linked together to form a (C₂-C₄)alkylene which is unsubstituted or substituted one or more timeswith a (C₁ -C₄)alkyl;

R₉ is hydrogen, a (C₁ -C₄)alkyl or a phenyl(C₁ -C₄)alkyl which isunsubstituted or substituted on the phenyl one or more times with R₁₁ ;

R₁₀ is a halogen, a (C₁ -C₄)alkyl, a (C₁ -C₄)alkoxy, a hydroxyl, anamino, a (C₁ -C₄)alkylamino or a di(C₁ -C₄) alkylamino;

R₁₁ is a halogen, a (C₁ -C₄)alkyl, a hydroxyl, a (C₁ -C₄).alkoxy or abenzyloxy;

R₁₂ is a halogen, a (C₁ -C₄)alkyl, a hydroxyl, a (C₁ -C₄)alkoxy, abenzyloxy, an oxo or a phenyl;

R₁₃ is a (C₁ -C₄)alkyl, a halogen or a hydroxyl;

R₁₄ is a methyl, an amino, a (C₁ -C₄)alkylamino, a di(C₁ -C₄)alkylamino,a tri(C₁ -C₄)alkylammonium, an amidino, a (C₁ -C₄)alkylamidino, aguanidino, a (C₁ -C₄)alkylguanidino, a hydroxyl, a (C₁ -C₄)alkoxy, a (C₁-C₄)alkoxycarbonyl, a group --AlkN(R₁₅)Alk'N(R₁₅)₂, or a heterocyclicradical chosen from. 1-pyrrolidinyl, 1-piperidyl, perhydro-1-azepinyl,pyridyl, imidazolyl, dihydroimidazolyl, imidazolidinyl, pyrimidinyl andindolyl;

R₁₅ and R'₁₅ are, independently of each other, hydrogen or a (C₁-C₄)alkyl;

R₁₆ is hydrogen;

R₁₇ is hydrogen;

Alk and Alk' are, independently of each other, a (C₁ -C₄)alkylene;

Z is a (C₂ -C₁₂)alkylene or a (C₁ -C₆)alkylene which is interrupted orsubstituted with a (C₅ -C₇)cycloalkyl or with a phenyl;

as well as the salts thereof with inorganic or organic acids.

Certain values for the substituents are preferred. Thus, the preferredcompounds of formula (I) are those which satisfy at least one of thefollowing conditions:

a --R₁ is a naphthyl, a quinolyl or a trichlorophenyl; R₂, R₃, R₄, R₅,R₆, R₇, R₈, R₉, R₁₆ and R₁₇ being as defined above for formula I;

b --R₂ is a phenyl which is unsubstituted or substituted with R₁₁ ; R₁,R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₆ and R₁₇ being as defined above forformula I;

c --NR₄ R₅ is a 1-pyrrolidinyl group; R₁, R₂, R₃, R₆, R₇, R₈, R₉, R₁₆and R₁₇ being as defined above for formula I;

d --C(NR₆)NR₇ R₈ is a 4,5-dihydro-2-imidazolyl; R₁, R₂, R₃, R₄, R₅, R₉,R₁₆ and R₁₇ being as defined above for formula I;

e --R₃ =R₉ =R₁₆ =R₁₇ =H; R₁, R₂, R₄, R₅, R₆, R₇ and R₈ being as definedabove for formula I;

and the salts thereof with inorganic or organic acids.

According to the invention, the preferred compounds are those offormula: ##STR6## in which: R_(2a) is a phenyl which is unsubstituted orsubstituted in a meta or para position with R₁₁ ; a 1-naphthyl or a2-naphthyl;

R₁, R₆, R₇, R₈ and R₁₁ are as defined above for (1);

and the salts thereof with inorganic or organic acids.

Most particularly, the preferred compounds are those of formula:##STR7## in which: R_(1a) is a 1-naphthyl, a 2-naphthyl, a2,4,6-trichlorophenyl or a 2-quinolyl;

R_(2a) is as defined above for (Ia);

and the salts thereof with inorganic or organic acids.

More particularly, the preferred compounds are those of formula (I'a) inwhich R_(2a) is a phenyl which is unsubstituted or substituted in a metaor para position with R₁₁.

Most particularly, the preferred compounds are those of formula (I),(Ia) or (I'a) having (R,R) isomerism on the C*-labelled carbon atoms.

The following abbreviations are used in the description and in theclaims:

Me: methyl

Et: ethyl

iPr: isopropyl

nBuOH: n-butanol

iPrOH: isopropanol

EtOH: ethanol

MeOH: methanol

Et₂ O: ether:diethyl ether

DMF: dimethylformamide

DCM: dichloromethane

THF: tetrahydrofuran

AcOH: acetic acid

EtOAc: ethyl acetate

DIPEA: diisopropylethylamine

DMAP: 4-dimethylaminopyridine

DCC: 1,3-dicyclohexylcarbodiimide

DCU: dicyclohexylurea

NSuOH: N-hydroxysuccinimide

NSu: succinimido

NBS: N-bromosuccinimide

Fmoc: fluorenylmethoxycarbonyl

Boc: tert-butoxycarbonyl

(Boc)₂ O: di-tert-butyl dicarbonate

Et₃ N: triethylamine

Bn: benzyl

Pd/C: palladium-on-charcoal

Sephadex® LH 20: sold by Pharmacia

Sephadex® G 25: sold by Pharmacia

Alcalase®: Carlsberg subtilisin sold by Novo (Denmark)

Penicillin amidase: penicillin amidohydrolase, sold by Sigma

BOP: benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate

K₂ CO₃ potassium carbonate

K₂ SO₄ : potassium sulphate

KHSO₄ : potassium hydrogen sulphate

KHSO₄ /K₂ SO₄ : solution of 16.66 g of KHSO₄ and 32.32 g of K₂ SO₄ in 1l of water

NaCl: sodium chloride

Na₂ SO₄ : sodium sulphate

MgSO₄ : magnesium sulphate

NaOH: sodium hydroxide

NH₄ OH: aqueous ammonia

HCl: hydrochloric acid

TFA: trifluoroacetic acid

hydrochloric ether: saturated solution of hydrogen chloride gas indiethyl ether

mPa.s: milliPascal/second

m.p.: melting point

RT: room temperature

NMR: nuclear magnetic resonance

DMSO: dimethyl sulphoxide

δ: chemical shift

s: singlet; bs: broad singlet; ds: doubled singlet; d: doublet; dd:doubled doublet; t: triplet; bt: broad triplet; q: quartet; quint:quintet; mt: multiplet; m: unresolved multiplet

The subject of the present invention is also the process for thepreparation of the compounds of formula (I) and the salts thereof. Thisprocess, referred to as process 1, is characterized in that:

a1) a compound of formula: ##STR8## in which R₁, R₂, R₃, R₄, R₅, R₉,R₁₆, R₁₇ and C* have the definitions given above for (I), in the form ofa pure enantiomer or a mixture of isomers in any proportion, is treatedwith an alcohol of formula R--OH in which R is a (C₁ -C₄)alkyl, inacidic medium, in order to form an intermediate imidate which is reactedwith an amine of. formula HNR₇ R₈ (III) or a diamine of formula H₂ NR₆R₈ NH₂ (IV) in which R₆, R₇ and R₈ have the definitions given above for(I);

b1) the compound of formula (I) thus obtained is isolated in base formor salt form,

c1) where appropriate, another salt of the compound of formula (I) isprepared.

Many processes for the synthesis of amidines are described in the book"The chemistry of amidines and imidates", D. G. Neilson, Ed. Saul Patai,Wiley and Sons, 1975, 389-394. The preparation of certain amidines isdescribed precisely in patent application EP 614,911 A.

The formation of the imidate is preferably carried out in a strong acidmedium, whereas the imido ester in free base form or in salt form isreacted with the amine (III) or the diamine (IV) in an inert polarsolvent, for example an alcohol, at a temperature of between 0° C. andthe reflux temperature of the solvent.

The intermediate imidate is reacted with an amine whose formula dependson that of the compound (I) which it is desired to obtain. In order toprepare a compound of formula (I) in which R₆ =H, an amine HNR₇ R₈ isreacted; in order to prepare a compound of formula (I) in which R₆ =R₈and R₇ =H, two moles of an amine of formula H₂ NR₈ are reacted per moleof imidate; in order to prepare a compound of formula (I) in which R₇ ishydrogen and R₆ and R₈ are linked together to form a (C₂ -C₄)alkylenewhich is unsubstituted or substituted one or more times with an alkyl, adiamine of formula H₂ NR₆ R₈ NH₂ is reacted.

Most of the amines (III) and of the diamines (IV) are known and thenovel products can be prepared by applying principles and methods thatare well known to those skilled in the art. For example, for thederivatives in which R₁₄ is an imidazolyl, reference will be made toU.S. Pat. No. 3,881,016 and to the publication Synth. Communic. 1987, 17(21), 223-227.

The compounds of formula (I) in which R₁₄ is NH₂ or alkylamino can beprepared by hydrolysis of the compounds of formula (I) in which R₁₄contains a t-butoxycarbonylamino group, which is in turn obtainedaccording to Synth. Commun. 1990, 20 (16), 2559-2564.

The compounds of formula (I) in which R₁₄ is a substituted orunsubstituted guanidino group can be prepared by the action on thecompound of formula (I), in which R₁₄ =NH₂, of a compound of formula:##STR9## in which T is H or (C₁ -C₄)alkyl and Y is a nucleofugal group,such as SO₃ H, for example, aminoiminomethanesulphonic acid (under theconditions described in Tetrahedron Letters, 1988, 3183-3186) orN-methylaminoiminomethanesulphonic acid (obtained according to theprocess described in J. Org. Chem., 1986, 51(10), 1882).

The action of an amine (III) of formula H₂ NR₈ in excess on the imidateresulting from the reaction of ROH with a compound of formula (II) leadsto the formation of a mixture of two compounds of formula (I): for oneR₆ =R₇ =H and for the other R₆ =R₈ and R₇ =H.

The compounds of formula (I) in which R₆ and R₈ together form a (C₂-C₄)alkylene which is unsubstituted or substituted one or more timeswith a (C₁ -C₄)alkyl can be prepared in a manner which is known per se,by the action of a diamine H₂ N--R₆ R₈ --NH₂ on the imido ester oroptionally by the action of the same diamine, one of the functions ofwhich is protected by a labile group (such as Boc or Fmoc for example)which will be removed before cyclization.

The compounds of formula (I) in which R₁₄ is a dihydroimidazole can beprepared by the action of an alcohol in acidic medium on a compound offormula: ##STR10## in order to form an intermediate imidate, which isreacted with an appropriate amine according to the usual methods.

The nitrites of formula (II) are prepared using the standard methods ofpeptide chemistry, for example those described in The Peptides Ed. E.Gross and J. Meienhofer, Academic Press, 1979, 1, 65-104. Known methodsmake it possible to carry out peptide couplings without racemization ofthe carbon atoms of each constituent amino acid; furthermore, theβ-substituted β-alanines for which the chiral carbon is not adjacent tothe carboxyl group are reputed as not suffering racemization (Ann. Rev.Biochem., 1986, 55, 855-878). Moreover, patent application EP 236,163describes processes which allow the chirality of each amino acid to beconserved.

In general, the coupling reactions between the 2 amino acids take placeat temperatures of between 0° C. and 40° C. in an inert solvent such asdichloromethane, acetonitrile, tetrahydrofuran or dimethylformamide, inthe presence of a coupling agent and of at least one equivalent of atertiary amine such as triethylamine, N-ethylmorpholine ordiisopropylethylamine.

Thus, the preparation of a nitrile of formula (II) can be carried outaccording to one of the synthetic routes below. ##STR11##

In order to prepare a compound of formula (V), which is useful in route1, the radical R₁ SO₂ is introduced in a conventional manner by theaction of a sulphonyl halide of formula: R₁ SO₂ Hal in which R₁ is asdefined above for (I) and Hal is a halogen, preferably chlorine, ona-compound of formula: ##STR12## in which R₂, R₃ and R₉ have themeanings given above for (I) and R' is hydrogen or a (C₁ -C₄)alkyl.

A compound of formula (V) in which R₃ is hydrogen can also be preparedin 2 steps : firstly, a compound of formula: ##STR13## in which Y is R₁SO₂ or a protecting group such as Boc or Fmoc, after which the carbonchain is extended by one atom using known methods, then, if necessary,the protecting group is removed and a sulphonyl halide of formula R₁ SO₂Hal is reacted and, lastly, the group R' is removed, if it is other thanH.

This procedure is suitable, for example, when R₂ and R₉ are linkedtogether and constitute a (C₃ -C₅)-alkylene which is unsubstituted orsubstituted with R₁₂ or a (C₂ -C₄)-alkylene which is interrupted by anoxygen atom or a sulphur atom which is unsubstituted or substituted oneor more times with R₁₂.

The sulphonyl halides are known or are prepared by known methods.

The compounds of formula (VI) are prepared by the action of an amineHNR₄ R₅ on an amino acid of formula: ##STR14## in which the amine isprotected, for example by a Boc or Fmoc group, followed by the removalof the protecting group.

In the reaction sequence of route 2, the amine is deprotected in acidicmedium (TFA) and the radical R₁ SO₂ is introduced in a conventionalmanner by the action of a sulphonyl halide of formula: R₁ SO₂ Ha'l inwhich R₁ is as defined above for (I) and Hal is a halogen, preferablychlorine.

In route 1, as in route 2, the group R₁ SO₂ is introduced in thepresence of a base, optionally in a two-phase medium, in the presence ofa phase-transfer catalyst.

A substituent R₉ can be introduced into a compound of formula (II) inwhich R₉ =H by known methods, for example by the action of a halide offormula R₉ Hal, in which Hal is a halogen atom, for example chlorine.

The compounds of formula (II) in which R₉ and R₁₆ together form amethylene group are prepared by the action of para-formaldehyde oncompounds of formula (II) in which R₉ =R₁₆ =H.

The compounds of formula (II) are novel and constitute a further aspectof the present invention.

Certain compounds of formula (V), which are toluenesulphonamide orphenylsulphonamide derivatives, have been described in the followingpublications:

Tetrahedron, 1993, 49 (48), 11329-11340;

Centralblatt, 1929, II, 1398;

J. Org. Chem., 1978, 43(23), 4438-4441;

J. Org. Chem., 1966, 31(7), 2385-2386.

Thus, the compounds of formula: ##STR15## in which R₁, R₂, R₃, R₉ and C*are as defined for (I); it being understood that:

when R₁ is phenyl or p-tolyl and R₃ is hydrogen, then R₂ and R₉,together with the carbon and nitrogen atoms to which they are attached,are not pyrrolidinyl or 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinolyl, and

when R₁ is p-tolyl and R₉ is hydrogen, then R₂ is not an unsubstitutedphenyl;

are novel and form part cf the invention.

In order to prepare a compound of formula (I) in which R₉ and R₁₁together form a methylene or ethylene group, a beta-amino acid offormula (XIII) ##STR16## in which R₉ and R₁₁ form a methylene orethylene group, is prepared using known methods, for example the onedescribed in J. Org. Chem., 1987, 52, 616-622.

In order to prepare a beta-amino acid of formula (XIII) in which R₃ andR₁₁ together form a methylene or ethylene group, known methods are used.Thus, methyl 1-aminoindane-2-carboxylate is prepared in 3 steps from1-indanone: according to J. Med. Chem., 1970, 650, the action of methylcarbonate in the presence of sodium hydride makes it possible to obtainmethyl (1-oxo)indane-2-carboxylate, then, according to J. Heterocycl.Chem., 1974, 11, 982, methyl 1-hydroxyiminoindan-2-carboxylate isprepared and, lastly, the hydroxylamine is reduced to amine in thepresence of a catalyst.

1-Amino-1,2,3,4--tetrahydronaphthalene-2-carboxylic acid can be preparedaccording to the method described in J. Chromatogr., A 1994, 676,297-302. Alternatively, according to another process, when the amidinegroup C(═NR₆)NR₇ R₈ contains no function liable to react in a subsequentstep under the peptide coupling conditions, an amino acid derivativecontaining the amidine group can be prepared from the correspondingderivative containing a cyano group, and the couplings required toobtain a compound according to the invention can then be carried out.

Thus, according to a further aspect, the subject of the presentinvention is another process for the preparation of a compound offormula (I), which is referred to as process 2, characterized in that:

a2) a compound of formula: ##STR17## in which X is hydrogen Dr a Bocgroup and R₄, R₅ and C* are as defined for (I), in the form of a pureenantiomer or a mixture of isomers in any proportion, is treated with analcohol of formula R--OH in which R is a (C₁ -C₄)alkyl, in acidicmedium, in order to form an intermediate imidate which is reacted withan amine of formula HNR₇ R₈ (III) or a diamine of formula H₂ NR₆ R₈ NH₂(IV) in which R₆, R₇ and R₈ have the definitions given above for (I):

b2) the compound thus obtained, of formula: ##STR18## is coupled eitherwith a compound of formula: ##STR19## in which R₂, R₃ and R₉ are asdefined for (I) and Pr is a protecting group, for example Boc or Fmoc,then, after deprotection of the amine in acidic medium, a sulphonylhalide of formula R₁ SO₂ Hal in which R₁ is as defined for (I) and Halis a halogen, for example chlorine, is reacted;

or with a compound of formula: ##STR20## in which R₁, R₂, R₃ and R₉ areas defined for (I): c2) the compound of formula (I) thus obtained isisolated in base form or in salt form;

d2) where appropriate, another salt of the compound of formula (I) isprepared.

The compounds of formulae (VI) and (XI) in optically pure form can beobtained from an ester, for example the racemic ethyl ester of4-cyanophenylalanine, according to the reaction scheme described below:

SCHEME 3 ##STR21##

The racemic ethyl ester of 4-cyanophenylalanine is described in patentapplication EP 614,911 A. The amine function of this compound isprotected in a conventional manner, by the action of (Boc)₂ O in thepresence of triethylamine. The action -of an enzyme, Alcalase®, on thecompound 4 thus obtained makes it possible to selectively hydrolyse theester function of the amino acid of (L) configuration and thus toisolate each of the compounds 5 and 6 in optically pure form (Synthesis,1983, 1041-1043).

The compound of (L) configuration is isolated in acid form: (L) 6. Thecompound of (D) configuration is isolated in the form of an ethyl ester:(D) 5; this compound is hydrolysed with sodium hydroxide in order toobtain the acid form: (D) 6. Each of these 2 compounds of formula 6 isthen treated with N-hydroxysuccinimide in an inert solvent such as DMFor dioxane, in the presence of a coupling agent such as DCC in order toobtain a compound of formula 7. The action of an amine HNR₄ R₅, followedby the action of trifluoroacetic acid, makes it possible to prepare thecompounds of formulae (L) (VI) and (D) (VI); the conventional reactionsdescribed above are then carried out in order to obtain the desiredamidines of formulae (L) (XI) and (D) (XI).

The beta-amino acids of formula XIII: ##STR22## or the correspondingaliphatic esters are known or can be prepared by various methods, forexample according to J. Am. Chem., 1936, 58, 299. One specific way ofpreparing the beta-amino acids consists in carrying out a chainextension starting with an alpha-amino acid, according to Tetrahedron,1994, 50, 9457-9470 or according to J. Chem. Soc., Perkin Transact. II,1977, 370.

More specifically, the preparation of certain beta-amino acids offormula (XIV) H₂ N--CH(R₂)--CH₂ --COOH is described in the followingpublications or patents.

    ______________________________________                                        R.sub.2         Reference                                                     ______________________________________                                         ##STR23##      (R, S): commercial  (R): Ber. 1910, 43, 2020  (S) J. Org.                     Chem., 1991, 56, 5883  J. Chem. Soc. Chem. Commun., 1993,                      1153                                                          ##STR24##      Heterocycles, 1989, 28, 1015  Bull. Soc. Chim. Fr., 1987,                     1079                                                           ##STR25##                                                                     ##STR26##                                                                     ##STR27##      Heterocycles, 1978, 1277-1285                                  ##STR28##      Tetrahedron, 1987, 43, 3509-3517                               ##STR29##      EP-355819                                                      ##STR30##      Tetrahedron Lett., 1988, 29, 6465                              ##STR31##      Heterocycles, 1989, 28, 1015                                   ##STR32##      J. Agric. Food Chem., 1977, 25, 965                            ##STR33##      Tetrahedron, 1994, 50 (31), 9457-9470  Tetrahedron Lett.,                     1990, 31, 5153-5156                                           ______________________________________                                    

The compounds (I) in which R₂ and R₉ are linked together and constitutea C₃ -C₅ alkylene which is unsubstituted or substituted with R₁₂ areprepared from compounds of formula (XIII) which are known or areprepared by known methods.

In order to obtain optically pure compounds of formula (XIII) or (XIV)it is possible, for example, to use a menthol ester according to thetechnique described in Tetrahedron Lett., 1988, 29, 6465-6466; a quinineor quinidine salt can also be used, according to Chem. Ber., 1910, 43,2020, or alternatively the phenylacetamido derivative of the compound offormula (XIV) in racemic form can be used for an enzymatic resolutionwith a penicillin amylase (Synlett, 1993, 339). Enantioselectivesyntheses can also be carried out: Tetrahedron, 1994, 50, 9517;Aldrichimica, Acta, 1994, 27(1), 3.

In order to prepare an alpha-hydroxy-beta-amino acid, the methoddescribed in Bull. Soc. Chim., France, 1940, 7, 593-603 can be used.

The affinity of the compounds according to the invention for thebradykinin B₁ receptors was measured on suspensions of MRC5 cellmembranes using a technique similar to that described by K. H. Schnecket al., in Eur. J. Pharmacol., 1994, 266, 227-282. In this test, theaffinity of [des-Arg⁹ ] bradykinin is between 10⁻⁶ M and 10⁻⁷ M, that of[des-Arg¹⁰ ]kallidin is 2×10⁻⁹ M; and the compounds of the inventionhave an affinity ranging down to 10⁻¹⁰ m.

The affinity of the compounds according to the invention for thebradykinin B₂ receptors was measured on suspensions of MRC5 cellmembranes according to a technique similar to that described by D. G.Sawutz et al., in Eur. J. Pharmacol., 1992, 227, 339-315. In that test,the affinity of bradykinin is close to 10⁻⁹ M and that of the compoundsof the invention varies around 10⁻⁶ M or 10⁻⁷ M.

The toxicity of the compounds according to the invention is compatiblewith their therapeutic use.

The compounds according to the invention may be useful for the treatmentor prevention of many pathologies, in particular inflammationpathologies and persistent or chronic inflammatory diseases (Drug Newsand Perspectives, 1994, 10(7), 603-611). By way of example, mention maybe made of:

neurogenic inflammation, pain (Brit. J. Pharmacol., 1993, 110, 193-198),septic shock, asthma, rheumatoid arthritis, inflammatory diseases of thejoints, burns (Pain, 1993, 53, 191-197), wounds, diseases of therespiratory tracts, for example rhinitis of viral or allergic origin,systematic inflammatory response syndrome, oedema (Brit. J. Pharmacol.,1995, 114, 1005-1013), angiogenesis (Brit. J. Pharmacol., 1993, 109,14-17), type-I infectious diabetes (Abst. 14th Intern. Symp. on Kinins,C49, Denver Colorado, Sep. 10-15, 1995), ventricular hypertrophyassociated with diabetes, pulmonary fibrosis and systemic progressivesclerosis, enterocolitis, and more generally any bradykinin-dependentpathology.

The compounds according to the invention are generally administered indosage units.

The invention also relates to pharmaceutical. compositions comprising,as active principle, one of the enantiomers of the compounds of formula(I), a mixture thereof or salts thereof with a pharmaceuticallyacceptable acid, as well as to an excipient which is suitable for oral,injectable, topical or transdermal administration. The daily dosesdepend on the pathology to be treated and on the patient.

The subject of the present invention is also pharmaceutical compositionscontaining an effective dose of a compound according to the invention orof a pharmaceutically acceptable salt and suitable excipients.

The said excipients are chosen depending on the pharmaceutical forme andthe desired mode of administration.

In the pharmaceutical compositions of the present invention for oral,sublingual, subcutaneous, intramuscular, intravenous, topical,intratracheal, intranasal, transdermal or rectal administration, theactive principles of formula (I) above, or the possible salts thereof,can be administered in unit forms of administration, as a mixture withstandard pharmaceutical supports, to animals and to human beings for theprophylaxis or treatment of the above disorders or diseases. Theappropriate unit forms of administration comprise oral forms such astablets, gelatin capsules, powders, granules or oral solutions orsuspensions, sublingual, buccal, intratracheal or intranasaladministration forms, subcutaneous, intramuscular or intravenousadministration forms and rectal administration forms. For topicalapplication, the compounds according to the invention can be used increams, ointments, gels cr lotions.

In order to obtain the desired prophylactic or therapeutic effect, thedose of active principle can range between 0.01 and 50) mg per kg ofbody weight and per day.

Each unit dose can contain from 0.5 to 1000 mg, preferably from 1 to 500mg, of active ingredients in combination with a pharmaceutical support.This unit dose can be administered 1 to 5 times a day so as toadminister a daily dose of from 0.5 to 5000 mg, preferably from 1 to2500 mg.

When a solid composition is prepared in the form of tablets, the mainactive ingredient is mixed with a pharmaceutical vehicle such asgelatin, starch, lactose, magnesium stearate, talc, gum arabic or thelike. The tablets can be coated with sucrose, with a cellulosederivative or with other suitable materials or alternatively they can betreated such that they have sustained or delayed activity and such thatthey release a predetermined amount of active principle continuously.

A preparation as gelatin capsules can be obtained by mixing the activeingredient with a diluent such as a glycol or a glycol ester and bypouring the mixture obtained into soft or hard gelatin capsules.

A preparation in syrup or elixir form or for administration in the formof drops can contain the active ingredient together with a sweetener,preferably a calorie-free sweetener, methylparaben and propylparaben asantiseptic, as well as a flavouring agent and a suitable dye.

The water-dispersible powders or granules can contain the activeingredient as a mixture with dispersing agents, wetting agents orsuspension agents, such as polyvinylpyrrolidone, as well as withsweeteners or flavour enhancers.

For rectal administration, use is made of suppositories which areprepared with binders which melt at the rectal temperature, for examplecocoa butter or polyethylene glycols.

Aqueous suspensions, isotonic saline solutions or sterile and injectablesolutions which contain pharmacologically compatible dispersing agentsand/or wetting agents, for example propylene glycol or butylene glycol,are used for parenteral administration.

Creams, ointments, lotions or gels, for example, can be used for localadministration.

Patches in multilaminar or reservoir form in which the active principlemay be dissolved can be used for transdermal administration.

The active principle can also be formulated in the form ofmicrocapsules, optionally with one or more supports or additives.

The compositions of the present invention can contain, along with theproducts of formula (I) above or one of the pharmaceutically acceptablesalts, other active principles which can be used in the treatment of thedisorders or diseases mentioned above.

The preparations and the examples below illustrate the invention. Exceptwhere otherwise mentioned, the compounds are obtained in the form of amixture of diastereoisomers.

The nuclear magnetic resonance (NMR) spectra are recorded at 200 MHz indeuterated DMSO optionally containing TFA, using tetramethylsilane asreference. The chemical shifts are indicated in ppm.

The mass spectra indicate the value MH⁺.

PREPARATIONS

Preparation 1.1

1-[2-Amino-3-(4-cyanophenyl)propionyl]pyrrolidine trifluoroacetate.

4.06 g of 2-[N-(Boc)amino]-3-(4-cyanophenyl)propionic acid are mixedwith 1.15 ml of pyrrolidine and 7.5 g of BOP in 20 ml of DMF; themixture is left stirring for 2 hours at RT while maintaining at pH 7 byaddition of Et₃ N. After evaporating to dryness, the residue ischromatographed on silica, eluting with a chloroform/methanol mixture(9/0.5; v/v). After evaporating off the solvents, the solid obtained istreated with 20 ml of TFA in 20 ml of DCM for 30 minutes at RT. Thesolvents are evaporated off and the residue is taken up in ether andthen dried in order to obtain 3.95 g of the expected product.

Preparation 1.2

1-[2-Amino-3-(4-(3,4-dihydroimidazol-2-yl)phenyl)propionyl]pyrrolidinedihydrochloride.

18 g of 1-[2-((N-Boc)amino)-3-(4-cyanophenyl)propionyl]pyrrolidine aredissolved in 90 ml of DCM, 90 ml of TFA are added and the mixture isleft stirring for 40 minutes at RT. After evaporating to dryness, theresidue is taken up in DCM and then evaporated (twice). The residue isthen taken up in 300 ml of HCl-saturated anhydrous ethanol at 0° C. andthe mixture is left for 18 hours at +4° C. The medium is evaporated todryness, taken up in ethanol, evaporated again and then taken up againin DCM and evaporated (twice). 23 g of intermediate imidate areobtained. The product obtained is dissolved in 1 liter of anhydrousethanol and 4.41 g of ethylenediamine in 50 ml of anhydrous ethanol areadded over 30 minutes. After stirring for 48 hours at RT, the medium isconcentrated and then acidified to pH=2 by addition of a saturatedsolution of HCl in methanol. The mixture is drained, evaporated todryness and then taken up in ether, drained and dried to give 17.4 g ofthe expected product.

NMR (DMSO+TFA) : 1.50-1.80 :m : 4H; 2.50-3.60 : m : 6H; 4.00 : s : 4H;4.35 : bs : 1H; 7.50 : d : 2H; 8.15 : d : 2H; 8.60 : bs : 3H; 11.10 : s: 2H.

Preparation 1.3

1-[2-Amino-3-(4-(N¹-[3-(dimethylamino)propyl]amidino)phenyl)propionyl]pyrrolidinetris(trifluoroacetate).

A) 1-[2-Amino-3-(4-N¹-[3-(dimethylamino)propyl]-amidino)phenyl)propionyl]pyrrolidinetrihydrochloride.

809 mg of 1-[2-amino-3-(4-cyanophenyl)propionyl]pyrrolidinetrifluoroacetate are dissolved in 50 ml of hydrochloric acid-saturatedmethanol at 0° C. and the mixture is left overnight in a refrigerator.After evaporating to dryness, the imidate formed is taken up in toluene,the mixture is then evaporated and the residue is dried under vacuumover potassium hydroxide. -The product obtained is dissolved in 75 ml ofanhydrous methanol and 570 μl of N-(dimethyl)propane-1,3-diaminedissolved in 15 ml of anhydrous methanol are added slowly. Afterstirring for 3 hours at RT, the mixture is evaporated to dryness and theresidue is then dissolved in 30 ml of methanol, adding 5 ml of 4N HCl indioxane, and is evaporated to dryness.

B) 1-[2-(N-Boc)amino-3-(4-(N¹-[3-(dimethylamino)propyl]amidino)phenyl)propionyl]pyrrolidinedihydrochloride.

The crude product obtained in the above step is dissolved in 10 ml ofdioxane and 10 ml of water. Triethylamine is added to reach pH=8.3,followed by 600 mg of (Boc)₂ O and the mixture is left stirring at RTfor 15 hours. The medium is diluted with water, washed twice with ether,acidified to pH=1.5 by addition of 1N HCl and then washed with DCM. Thismixture is brought to pH=7 by addition of 1N sodium hydroxide and isevaporated to dryness. The residue is taken up in DCM, the insolublematerial is filtered off and the filtrate is then purified bychromatography on silica, eluting with a chloroform/methanol mixture(85/15 to 80/20; v/v). 560 mg of the expected product are obtained.

C) 1-[2-Amino-3-(4-(N¹-[3-(dimethylamino)propyl]-amidino)phenyl)propionyl]pyrrolidinetris(trifluoroacetate).

550 mg of the compound of step B are mixed with 15 ml of DCM and 15 mlof TFA and the mixture is left stirring for 45 minutes at RT. Afterevaporating to dryness, the residue is dissolved in isopropanol,evaporated to dryness again and taken up twice in ether. The product isisolated by decanting and then dried under vacuum in the presence ofpotassium hydroxide. 380 mg of the expected compound are obtained.

Preparation 1.4

1-[2-Amino-3-(4-cyanophenyl)propionyl]-2,5-dimethyl-2,5-dihydropyrroletrifluoroacetate.

A)1-[2-(N-Boc)amino-3-(4-cyanophenyl)propionyl]-2,5-dimethyl-2,5-dihydropyrrole.

2.1 g of 2-[(N-Boc)amino]-3-(4-cyanophenyl)propionic acid are mixed with0.715 g of 2,5-dimethyl-2,5-dihydropyrrole in 10 ml of DMF, 3.9 g of BOPare added and the pH is adjusted to 7 by addition of Et₃ N. Afterstirring for 18 hours at RT, the mixture is evaporated to dryness and isthen taken up in EtOAc, washed with NaHCO₃ solution, with KHSO₄ /K₂ SO₄and with saturated NaCl solution,, The resulting solution is dried overNa₂ SO₄ and then evaporated. The residue is taken up in an ether/hexanemixture and then dried over Na₂ SO₄. 1.4 g of the expected compound areobtained.

NMR : (DMSO+TFA) 1.15-1.25 : m : 6H; 1.30 : s : 9H; 2.9-3.1 : m : 2H;4.3-4.6 : m : 1H; 4.6-4.75 : m : 1H; 4.9-5.1 : m : 1H; 5.75-5.9 : m :2H; 7.5 : d : 2H; 7.9 : d : 2H.

B)1-[2-Amino-3-(4-cyanophenyl)propionyl]-2,5-dimethyl-2,5-dihydropyrroletrifluoroacetate.

The compound of the above step is dissolved in 5 ml of DCM, 5 ml of TFAare added and the mixture is left stirring for 40 minutes at RT. It isevaporated to dryness and the residue is taken up in DCM and evaporated(3 times) and is then taken up in an ether/hexane mixture. The productis drained and then dried over Na₂ SO₄ in order to obtain 1.5 g of theexpected product.

Working as in Preparation 1.4, step A above, the compounds described inthe table below are prepared:

                  TABLE 1                                                         ______________________________________                                         ##STR34##                                                                    Preparation                                                                           --NR.sub.4 R.sub.5                                                                        NMR (DMSO + TFA)                                          ______________________________________                                        1.5                                                                                    ##STR35##  1.20: s: 9H; 2.75-2.95: mt: 2H;  3.30-3.60: m: 8H;                            4.60: mt: 1H; 7.40:  d: 2H; 7.70: d: 2H                   1.6                                                                                    ##STR36##  0.8-1.2: m: 6H; 1.3: s: 9H; 2.7-3: m:  5H; 4-4.8: m:                          2H; 7.5: d: 2H; 7.8:  d: 2H                               1.7                                                                                    ##STR37##  0.8-1.1: m: 6H; 1.15: 2: 9H; 1.5-2.1:  m: 4H;                                 2.7-2.9: m: 2H; 3.6-4: m: 2H;  4.1-4.4: m: 1H; 7.4:                           d: 2H; 7.7: d: 2H                                         ______________________________________                                    

Preparation 1.8

1-[2-N-Methylamino-3-(4-cyanophenyl)propionyl]-pyrrolidinetrifluoroacetate.

A) 2-(N-Boc-N-methyl)amino-3-(4-cyanophenyl)propionic acid.

1.16 g of 2-[N-Boc-amino]-3-(4-cyanophenyl)propionic acid are dissolvedin 20 ml of THF and 2 ml of methyl iodide are added at 0° C., followedby portionwise addition of 360 mg of sodium hydride at 80% in oil. Themixture is left stirring overnight at RT. The reaction medium is dilutedwith EtOAc and then water is added and the pH is brought to 2.5 with 1NHCl. The organic phase is separated out after settling has taken place,then washed with water, with saturated NaCl solution and then dried overNa₂ SO₄ and evaporated. The residue is taken up in an Et₂ O/hexanemixture (1/1; v/v). The powder formed is filtered off and dried to give1.11 g of the expected compound.

NMR (DMSO+TFA) : 1.20 : ds : 9H; 2.55 : ds : 3H; 2.90-3.25 : m : 2H;4.50-4.80 : m : 1H; 7.30-7.70 : m : 4H.

B) 1-[2-(N-Boc-N-methyl)amino-3-(4-cyanophenyl)propionyl]pyrrolidine.

1.10 g of the compound of the above step, 0.35 ml of pyrrolidine and1.78 g of BOP-are mixed with stirring in 15 ml of DMF and the pH isadjusted to 6 by addition of DIPEA. After stirring for 2 and a halfhours, the mixture is extracted with EtOAc and the organic phase is thenwashed with 0.25N NaOH, 0.25N HCl, H₂ O and then with saturated NaClsolution. The thick wax formed sets to a solid after a few days at +4°C. 1.25 g of the expected compound are obtained.

NMR (DMSO+TFA) : 1.10 : ds : 9H; 1.60-1.85 : m : 4H; 2.60 : ds : 3H;2.75-3.40 : m : 6H; 4.80-5.10 : m : 1H; 7.25-7.70 m : 4H.

C) 1-[2-N-Methylamino-3-(4-cyanophenyl)propionyl]-pyrrolidinetrifluoroacetate.

0.72 g of the compound of the above step is placed in 12 ml of TFA and12 ml of DCM. After stirring for 40 minutes at RT, the reaction mediumis concentrated under vacuum and then evaporated with DCM. 0.75 g of theexpected compound is obtained in the form of a thick-wax.

Preparation 1.9

N,N-Diethyl-[2-amino-3-(4-(3,4-dihydroimidazol-2-yl)phenyl)]propionamidedihydrochloride.

N,N-Diethyl-[2-(N-Boc)amino-3-(4-cyanophenyl)]-propionamide is preparedby working according to the procedure described in Preparation 1.4, stepA. The expected product is then obtained according to the proceduredescribed in Preparation 1.2.

Preparation 1.10

Ethyl ester of 2-(N-Boc)amino-3-(4-cyanophenyl)propionic acid.

26 g of Boc₂ O dissolved in 100 ml of DCM are gradually added to asolution of 25.5 g of the ethyl ester hydrochloride of2-amino-3-(4-cyanophenyl)propionic acid and 13.9 ml of Et₃ N in 400 mlof DCM. After stirring for 6 hours at RT, the reaction medium is washedwith KHSO₄ /K₂ SO₄ solution, with saturated NaHCO₃ solution and withsaturated NaCl solution. After drying over Na₂ SO₄ and evaporation ofthe DCM, the residue is triturated from heptane to give 29 g of whitepowder.

Preparation 1.11

Ethyl ester of (R)-2-(N-Boc)amino-3-(4-cyanophenyl)propionic acid :compound B and (S)-2-(N-Boc)amino-3-(4-cyanophenyl)propionic acid :compound A.

A mixture of 24 g of the product obtained above and 8.4 g of NaHCO₃ in900 ml of EtOAc and 500 ml of H₂ O is treated with 2 ml of Alcalase® for24 hours at RT. The 2 phases are separated out after settling has takenplace; the EtOAc phase is rewashed with 100 ml of 10% NaHCO₃ solutionwhich is added to the first aqueous phase and the aqueous phase isrewashed with 100 ml of EtOAc added to the firs: EtOAc phase. The EtOAcphase thus obtained is dried over Na₂ SO₄ and then evaporated todryness; 12.45 g of compound B are obtained.

α_(D) ²⁵ =+8.8° (c=1; MeOH)

The aqueous phase is taken up in EtOAc and brought to pH=2.5 with 6NHCl. EtOAc is separated out after settling has taken place and rewashedwith KHSO₄ /K₂ SO₄ solution, with saturated NaCl solution, dried overNa₂ SO₄ and evaporated to dryness, and 10.1 g of compound A areobtained.

α_(D) ²⁵ =+8.8° (c=1; MeOH)

Preparation 1.12

1-[2-Amino-3-(4-cyanophenyl)propionyl]pyrrolidine trifluoroacetate, (S)isomer.

A) 2,5-Dioxo-1-pyrrolidinyl ester of2-(N-Boc)amino-3-(4-cyanophenyl)propionic acid, (S) isomer.

9.85 g of 2-(N-Boc)amino-3-(4-cyanophenyl)propionic acid, (S) isomer,and 4.14 g of NSuOH are dissolved in 100 ml of dioxane and 8.5 g of DCCdissolved in 30 ml of dioxane are added slowly at RT, after which themixture is stirred for 8 hours at RT. The DCU is filtered off and washedwith acetone. After evaporating the filtrate to dryness, the residue isdissolved in acetone and then left overnight at RT. The remaining DCUwhich has precipitated is removed and the filtrate is then evaporated todryness, triturated from Et₂ O and then drained, washed with Et₂ O anddried. 11.5 g of the expected compound are obtained,

α_(D) ²⁵ =-29.7° (c=1, MeOH)

B) 1-[2-(N-Boc)amino-3-(4-cyanophenyl)propionyl]-pyrrolidine, (S)isomer.

11 g of the compound of the above step are placed in 150 ml ofacetonitrile and 20 ml of DMF and 2.5 ml of pyrrolidine in 30 ml ofacetonitrile are added over 10 minutes. The mixture is left stirring for2 hours at RT and is then left overnight at RT. It is evaporated todryness, the residue is taken up in EtOAc and a KHSO₄ /K₂ SO₄ buffer isthen added. This mixture is extracted with EtOAc, then washed withsaturated NaHCO₃ solution and then with saturated NaCl solution; it isdried over Na₂ SO₄ and then evaporated to dryness. The residue istriturated from Et₂ O, drained, washed with Et₂ O and dried. 6.95 g ofthe expected compound are obtained.

α_(D) ²⁵ =+26° (c=1, MeOH)

C) 1-[2-Amino-3-(4-cyanophenyl)propionyl]pyrrolidine trifluoroacetate,(S) isomer.

6.75 g of the compound of the above step are dissolved in 50 ml of DCMand the insoluble material is filtered off. 50 ml of TFA are added andthe mixture is left stirring for 45 minutes. It is evaporated todryness, the residue is redissolved in isopropanol, this solution isagain evaporated to dryness and the residue is then triturated from Et₂O, drained, washed with Et₂ O and dried over Na₂ SO₄. 6.13 g of theexpected compound are obtained, m.p.=193-196° C.

α_(D) ²⁵ =+51° (c=1, MeOH)

Preparation 1.13

1-[2-Amino-3-(4-cyanophenyl)propionyl]pyrrolidine trifluoroacetate, (R)isomer.

A) (R)-2-(N-Boc)amino-3-(4-cyanophenyl)propionic acid.

43 ml of 1N NaOH solution are added to 12.18 g of compound B ofPreparation, 1.11 dissolved in 180 ml of MeOH and the mixture is stirredfor 1 hour at RT. 43 ml of 1N HCl solution are then added and 150 ml ofmethanol are evaporated off, after which the mixture is taken up inEtOAc and is washed with water and then with saturated NaCl solution. 11g of the expected compound are obtained after crystallization from anEt₂ O/heptane mixture.

α_(D) ²⁵ =-9.5° (c=1; MeOH)

B) Ester of 2,5-dioxo-1-pyrrolidinyl(R)-2-(N-Boc)amino-3-(4-cyanophenyl)propionic acid.

4.2 g of NSuOH are added to 10 g of the acid obtained above dissolved in10 ml of dioxane, followed by addition, over 20 minutes, of 8.62 g ofDCC dissolved in 30 ml of dioxane. After stirring overnight at RT, theDCU formed is filtered off and washed with dioxane. The filtrate isevaporated to dryness and the residue is triturated from ether to give asolid which is filtered off and dried. 12.09 g of the expected compoundare obtained.

α_(D) ²⁵ =+27.1° (c=1; MeOH)

C) 1-[2-(N-Boc)amino-3-(4-cyanophenyl)propionyl]-pyrrolidine, (R)isomer.

2.6 ml of pyrrolidine dissolved in 20 ml of acetonitrile are added to11.6 g of the compound obtained in the above step dissolved in 150 ml ofacetonitrile plus 20 ml of DMF. After stirring overnight at RT, a smallamount of insoluble material is removed and the filtrate is concentratedunder vacuum. The residue is taken up in EtOAc and washed with KHSO₄ /K₂SO₄ solution, with saturated NaHCO₃ solution and with saturated NaClsolution; after drying over Na₂ SO₄, the EtOAc is evaporated undervacuum, the residue is triturated from ether and 9.3 g of the expectedcompound are obtained in the form of a white solid.

α_(D) ²⁵ =-29.2° (c=1; MeOH)

D) 1-[2-Amino-3-(4-cyanophenyl)propionyl]pyrrolidine trifluoroacetate,(R) isomer.

8.7 g of the product obtained in the above step are stirred for 35minutes in a mixture of 50 ml of DCM and 50 ml of TFA. After evaporatingto dryness, the residue is taken up in isopropanol and re-evaporated todryness, and 8.67 g of the expected compound are obtained in solid form.

α_(D) ²⁵ =-46° (c=1; MeOH)

Preparation 1.14

1-[2-Amino-2-methyl-3-(4-cyanophenyl)propionyl]-pyrrolidinetrifluoroacetate.

A)

A mixture of 7 g of (D,L) alanine methyl ester, 14 ml of Et₃ N, 4.2 g ofMgSO₄.3H₂ O and 5.1 ml of benzaldehyde in 100 ml of dichloromethane isstirred for 18 hours at RT. The insoluble material is filtered off, thefiltrate is concentrated under vacuum and the residue is taken up in anether/water mixture; the ether is separated out after settling of thephases has taken place, rewashed with water and then with saturated NaClsolution, dried over Na₂ SO₄ and evaporated. 8.3 g of the expectedcompound are obtained in the form of an oil.

B)

13.2 ml of lithium bis(trimethylsilyl)amide (IM in THF) are added, at-70° C. over 20 minutes, to 2.29 g of the product of the above stepdissolved in 60 ml of THF. After 30 minutes, 2.35 g of4-bromomethylbenzonitrile dissolved in 15 ml of THF are added over 15minutes and the temperature is then allowed to rise slowly. After 3 anda half hours, the reaction medium is taken up in EtOAc and washed withwater and with saturated NaCl solution. After drying and evaporating theEtOAc, 3.6 g of the expected compound are obtained in the form of anoil.

C)

The compound of the above step is taken up in 60 ml of ether plus 60 mlof 1N HCl and is stirred for 18 hours at RT. The aqueous phase isseparated out after settling of the phases has taken place, placed incontact with EtOAc and brought to pH=10 with 10N NaOH; the EtOAc isseparated out after settling of the phases has taken place, rewashedwith H₂ O and with saturated NaCl solution, dried and evaporated. 2.20 gof the expected compound are obtained in the form of an oil.

D)

2.42 g of Boc₂ O dissolved in 10 ml of dioxane are added, at 10° C. over10 minutes, to 2.18 g of the compound of the above step dissolved in 10ml of dioxane; the mixture is then stirred overnight at RT and then for3 hours at 40° C. The reaction medium is taken up in EtOAc, washed withwater and then with saturated NaCl solution, dried and then evaporatedto give an oil which is used directly in the following step.

E)

The product of the above step is dissolved in 10 ml of methanol, 2.4 mlof 8.36N KOH solution are added and the mixture is stirred overnight; afurther 2.4 ml of the KOH solution are added and the mixture is refluxedfor 1 hour. The reaction medium is cooled and taken up in a water/ethermixture; the aqueous phase is separated out after settling of the phaseshas taken place, taken up in EtOAc and brought to pH=2 with 1N HCl. TheEtOAc is separated out after settling of the phases has taken place,rewashed with H₂ O and with saturated NaCl solution, dried over Na₂ SO₄and then evaporated. 0.99 g of the expected compound is obtained afterchromatography on silica, eluting with chloroform/MeOH/AcOH (94/6/0.2;v/v/v).

F)

0.30 ml of pyrrolidine, 1.55 g of BOP and DIPEA to obtain a pH of 6-7are added to 0.98 g of the compound of the above step in 10 ml of DMF.After stirring for 2 and a half hours, the reaction medium is taken upin EtOAc and washed with H₂ O, 0.2N NaOH, 0.2N HCl, H₂ O and saturatedNaCl solution. After drying and evaporating, 1.13 g of the expectedcompound are obtained, m.p.=84-87° C.

NMR (DMSO+TFA) : 1.20 : s : 3H; 1.45 : s : 9H; 1.60-1.90 : m : 4H;3.00-3.50 : m : 6H; 7.30 : d : 2H; 7.80 : d : 2H.

G)

1.12 g of the compound of the above step are stirred for 1 hour in amixture of 10 ml of DCM and 12 ml of TFA. After evaporating and drying,1.15 g of the expected compound are obtained in the form of a solid.

Preparation 2.1

3-Amino-3-(2-naphthyl)propionic acid hydrochloride.

The method described in J. Am. Chem. Soc., 1936, 58, 299 is used. 15.5 gof 2-formylnaphthalene, 10.4 g of malonic acid and 15.4 g of ammoniumacetate in 150 ml of ethanol are mixed together and refluxed for 6hours. After cooling to RT, the product is drained, washed with EtOH anddried. The product obtained is dissolved in a sufficient amount of 2NHCl and the insoluble material is then filtered off. The acidic solutionis concentrated by evaporating under vacuum and the solid thus formed isrecrystallized from 20 ml of an AcOH/H₂ O mixture (1/1; v/v). 3.8 g ofthe expected product are obtained.

The compounds of Table 2 below are prepared according to the proceduredescribed above.

                  TABLE 2                                                         ______________________________________                                         ##STR38##                                                                    Preparations                                                                              R.sub.2                                                           ______________________________________                                        2.2                                                                                        ##STR39##                                                        2.3                                                                                        ##STR40##                                                        2.4                                                                                        ##STR41##                                                        2.5                                                                                        ##STR42##                                                        ______________________________________                                    

Preparation 2.6

3-(N-Boc)amino-3-(3,4-dichlorophenyl)propionic acid.

Starting with the compound of Preparation 2.5, the N-protectedbeta-amino acid is prepared in the following way.

10 g of 3-amino-3-(3,4-dichlorophenyl)propionic acid are dissolved in100 ml of water and 9 ml of triethylamine are added, followed by gradualaddition of 9.3 g of (Boc)₂ O in 100 ml of dioxane. After stirringovernight at RT, the medium is concentrated and the residue is thentaken up in water, adjusting the pH to 9 by addition of 1N sodiumhydroxide. This mixture is washed twice with ether, the aqueous phase istaken up and acidified to pH=3 by addition of 1N HCl and is extractedwith EtOAc and concentrated in order to obtain 3.9 g of the expectedcompound, m.p.=128° C.

NMR (DMSO) 1.25 ppm : s : 9H; 2.50-2.65 ppm : mt : 2H; 4.75 ppm : q :1H; 7.25 ppm : d : 1H; 7.20-7.40 ppm : m : 3H

Preparation 2.7

3-Amino-3-(3-isopropyloxyphenyl)propionic acid hydrochloride

A) (3-Isopropyloxy)benzaldehyde.

3.4 g of K₂ CO₃ and 1.5 g of benzyltriethylammonium chloride are addedto 12.2 g of 3-hydroxybenzaldehyde in 100 ml of DMSO, followed byaddition, over 30 minutes, of 10 ml of isopropyl iodide dissolved in 20ml of DMSO, and the mixture is left stirring overnight at RT. Themixture is poured onto 400 ml of water and is then extracted with EtOAc,washed with saturated NaCl solution and then dried over Na₂ SO₄ andevaporated to dryness. The oil obtained (14.4 g) is used without furtherpurification in the following step.

B) 3-Amino-3-(3-isopropyloxyphenyl)propionic acid hydrochloride.

A mixture containing 14 g of the oil obtained in the above step, 150 mlof methoxyethanol, 8.95 g of malonic acid and 13.2 g of ammonium acetateis heated overnight at 80° C. After cooling, the mixture is evaporatedto dryness and the oil formed is then dissolved in ethanol and 50 ml of2N HCl. Fractional evaporation is carried out in order to obtain theexpected compound, which is drained and then washed with EtOAc. 4.56 gare obtained.

NMR (DMSO) : 1.25 : d : 6H; 4.65 : septet : 1H; 7.20-7.55 : mt : 4H;9.95 : s : 1H.

The compounds described in the table below are prepared according to theprocedure described in the above preparation.

                  TABLE 3                                                         ______________________________________                                         ##STR43##                                                                    Preparation                                                                           R.sub.2         NMR                                                   ______________________________________                                        2.8                                                                                    ##STR44##      2.85-3.15: mt: 2H; 3.85: s:  3H; 4.60: t: 1H;                                 6.95-7.45:  mt: 4H                                    2.9                                                                                    ##STR45##      (DMSO + TFA)  2.50-3.00: mt: 2H; 3.70: s:  3H;                                4.50: t: 1H; 6.90: d: 2H;  7.40: d: 2H                2.10                                                                                   ##STR46##      (DMSO + TFA)  2.80-3.10: mt: 2H; 4.60: t:  1H;                                7.30-7.60: m: 4H                                      2.11                                                                                   ##STR47##      (DMSO + TFA)  2.90-3.20: mt: 2H; 4.75: t:  1H;                                7.00-8.05: mt: 4H                                     2.12                                                                                   ##STR48##      (DMSO + TFA)  3.00-3.20: mt: 2H; 5.40-5.60:  mt:                              1H; 7.50-8.30: m: 7H                                  2.13                                                                                   ##STR49##      (DMSO + TFA)  2.75-3.10: mt: 2H; 5.60: t:  1H;                                7.00-7.45: m: 4H                                      2.14                                                                                   ##STR50##      (DMSO + TFA)  0.90-1.75: m: 11H;  2.40-2.70: mt:                              2H;  3.15-3.30: m: 1H                                 2.15                                                                                   ##STR51##      (DMSO + TFA)  2.25: s: 3H; 2.80-3.05: mt:  2H;                                4.55: t: 1H; 7.10-7.30:  m: 4H                        ______________________________________                                    

Preparation 2.16

3-(N-Boc)amino-3-biphenyl-4-ylpropionic acid.

A) 3-Amino-3-biphenyl-4-ylpropionic acid hydrochloride.

18.2 g of 4-phenylbenzaldehyde, 10.4 g of malonic acid and 15.4 g ofammonium acetate in 150 ml of methoxyethanol are mixed together. Afterheating overnight at 80° C., the mixture is cooled and the productformed is then washed with ethanol, with ether and then dried. Afterwashing again with water, the product is recrystallized from amethanol/water mixture with a small amount of HCl. A mixture of theexpected product and of the methyl ester hydrochloride of3-amino-3-biphenyl-4-ylpropionic acid is obtained, which is used withoutfurther purification in the following step.

B) 3-(N-Boc).Amino-3-biphenyl-4-ylpropionic acid.

The product of the above step is placed in 200 ml of dioxane, 55 ml of2N NaOH are added and the mixture is left stirring at RT for 1 hour 40minutes. 13 g of Boc₂ O are added and stirring is continued overnight.The insoluble material is filtered off, dilution is carried out with 200ml of water, the mixture is washed with Et₂ O and then acidified topH=2.5 by addition of 2N HCl in the presence of 100 ml of EtOAc. Thismixture is extracted with EtOAc, washed with KHSO₄ /K₂ SO₄, withsaturated NaCl solution and then dried over Na₂ SO₄ and evaporated todryness in order to obtain 11.03 g of the expected compound.

NMR (DMSO) : 1.40 : s : 9H; 2.60-2.85 : mt : 2H; 5.00 : dq : 1H;7.35-7.80 : mt : 10H; 12.30 : bs : 1H.

Preparation 2.17

2,5-Dioxo-1-pyrrolidinyl 3-(N-Boc)amino-3-biphenyl-4-ylpropionate.

3.41 g of the acid described in the above preparation are placed in 50ml of dioxane and treated with 1.26 g of hydroxysuccinimide in thepresence of 2.5 g of DCC. The mixture is left stirring overnight at RTand is then drained and washed with acetone. The filtrate is evaporatedto dryness and then taken up in isopropanol. The solid is drained,washed with ether and dried to give 3.46 g of the expected compound,m.p.=161° C.

NMR (DMSO) : 1.35 : s : 9H; 2.80 : s : 4H; 3.00-3.20 : mt : 2H; 5.00 :dq : 1H; 7.30-7.75 : mt : 10H.

Preparation 2.18

(1,2,3,4-Tetrahydro-1-isoquinolyl)acetic acid.

This compound is prepared according to J. Org. Chem., 1987, 52, 616-622.

A) 3,4-Dihydroisoquinoline.

9.4 ml of 1,2,3,4-tetrahydroisoquinoline are dissolved in 200 ml of DCMand 14.7 g of NBS are gradually added. The mixture is left stirring atRT, cooling slightly in order to maintain a temperature of less than 40°C. for 30 minutes, after which 50 ml of 10N NaOH are added and themixture is left stirring for 1 hour at RT. The organic phase isseparated out after settling of the phases has taken place and is washedwith 100 ml of water and then twice with 100 ml of 4N HCl. The aqueousphases are washed with DCM and then brought to pH=9 by addition ofconcentrated NH₄ OH. This mixture is extracted with DCM, dried over Na₂SO₄ and evaporated to dryness. The oil obtained is distilled off. 7.16 gof the expected compound are obtained, b.p.=50-54° C. at 0.05 mbar.

B) (1,2,3,4-Tetrahydro-1-isoquinolyl)acetic acid.

A mixture containing 6.62 g of 3,4-dihydroisoquinoline and 5.13 ofmalonic acid is triturated in an oil bath at 120° C. The mixturethickens and becomes entirely solid; it is worked into a powder with aspatula. The total duration of heating is about 40 minutes. The mixtureis cooled to about 60° C. and treated with 120 ml of MeOH and 20 ml ofwater, which dissolves the medium. A first crop is obtained under coldconditions and a second crop is then obtained by evaporating thefiltrate and crystallizing the residue from acetone.

NMR (DMSO+TFA) : 2.90-3.15 : mt : 4H; 3.30-3.60 : mt : 2H; 4.90 : t :1H; 7.20-7.40 : m : 4H.

Preparation 2.19

3-Amino-3-phenylpropionic acid trifluoroacetate, (R) isomer.

A) (2-Isopropyl-5-methyl)cyclohexyl 3-(N-Boc)amino-3-phenylpropionate,(R) isomer.

This reaction is carried out according to Tetrahedron Letters, 1988, 29,6465-6466. A solution containing 11.1 g of3-(N-Boc)amino-3-phenylpropionic acid, 7.5 g of L(-)-menthol and 2.1 gof DMAP in 400 ml of DCM is prepared; after stirring, 11.2 g of DCCdissolved in 50 ml of DCM are gradually introduced. After stirringovernight at RT, the mixture is filtered, the DCU is washed with acetoneand the filtrate is evaporated to dryness. The residue is taken up in150 ml of heptane at 80-90° C.; the insoluble material is filtered offand the solution is left at RT for 4 hours. The solid is drained, washedwith heptane and dried at 40° C. until the menthol odour hasdisappeared. 4.25 g of the expected compound are obtained.

α_(D) ²⁵ =-16.5° (c=1; MeOH)

B) 3-(N-Boc)amino-3-phenylpropionic acid, (R) isomer.

A mixture-containing 4.22 g of the compound of the above step and 15.7ml of 1N NaOH in 100 ml of methanol is refluxed for 4 hours. Thereaction medium is cooled, treated with 15.7 ml of 1N HCl and thenevaporated to dryness and taken up in heptane. After leaving for a fewhours at RT, the product crystallizes, it is drained, washed withheptane and then dried at 40° C. until the menthol odour hasdisappeared. 2.65 g of the expected compound are obtained.

α_(D) ²⁵ =+42.1° (c=1; MeOH)

C) 3-Amino-3-phenylpropionic acid trifluoroacetate, (R) isomer.

2.3 g of the compound obtained in the above step are dissolved in 15 mlof DCM and 15 ml of TFA are added. After stirring for 35 minutes at RT,the solution is evaporated to dryness. The product is taken up inisopropanol, evaporated and then crystallized from Et₂ O. The-product isdrained, washed with Et₂ O and dried in order to obtain 2.16 g of theexpected compound.

Preparation 2.20

3-Amino-4-phenylbutyric acid hydrochloride, (S) isomer.

A) Methyl 2-(N-Boc)amino-3-phenylpropionate, (S) isomer.

10.8 g of methyl (L)-2-amino-3-phenylpropionate hydrochloride in 150 mlof DCM are mixed with 7 ml of Et₃ N and 13 g of (Boc)₂ O in 50 ml of DCMare gradually added. After stirring for 5 hours at RT, the reactionmedium is washed with KHSO₄ /K₂ SO₄ and then with saturated NaClsolution. The resulting solution is dried in order to remove the excess(Boc)₂ O and is then dissolved in DCM and 1.5 ml ofN,N-dimethylpropanediamine are added. After stirring for 4 hours, themixture is washed with KHSO₄ /K₂ SO₄ and then with saturated NaClsolution. This solution is dried over Na₂ SO₄ and then evaporated todryness in order to obtain 11.9 g of the expected compound.

B) 2-(N-Boc)amino-3-phenylpropanol, (S) isomer.

This step and the following 2 are carried out according to Tetrahedron,1994, 50(31), 9457. 10 g of the compound of the above step are dissolvedin 120 ml of THF and the solution is cooled on an ice bath; 3.17 g oflithium chloride are added, followed by 2.8 g of sodium borohydride and,gradually, 170 ml of EtOH. After stirring overnight at RT, 70 ml of 1MKHSO₄ are added slowly and the mixture is concentrated almost todryness. The concentrate is diluted in chloroform and 1M KHSO₄ and isthen extracted with chloroform. The extracts are washed with saturatedNaCl solution, dried over Na₂ SO₄ and evaporated to dryness in order toobtain 8.65 g of the expected compound.

C) 2-Amino-3-phenylpropyl methanesulphonate, (S) isomer.

7.5 g of the compound of the above step are dissolved in 40 ml ofpyridine and the mixture is cooled in an ice bath; 3.3 ml of mesylchloride are added over 15 min and the mixture is then left stirring for2 hours at RT. 15 ml of water are added over 5 minutes and the medium isthen diluted with ether and washed with 1M KHSO₄ (twice), with water andthen with saturated NaCl solution; the resulting solution is dried overNa₂ SO₄ and then evaporated to dryness in order to obtain 9.2 g of theexpected compound.

α_(D) ²⁵ =-24.3° (c1; MeOH)

D) 3-(N-Boc)amino-4-phenylbutyronitrile, (S) isomer.

A solution containing 9.05 g of the compound of the above step with 7.3g of 18-crown-6 crown ether and 120 ml of DMSO is prepared and 9 g ofpotassium cyanide are added with stirring in an ice bath. After heatingat 50° C. for 5 hours, the mixture is cooled and 600 ml of Et₂ O arethen added. This mixture is washed with water (3 times) and then withsaturated NaCl solution; the resulting solution is dried over Na₂ SO₄and then evaporated to dryness in order to obtain 6.58 g of the expectedcompound.

E) 3-Amino-4-phenylbutyric acid hydrochloride, (S) isomer.

This step is carried out according to Tetrahedron Letters, 1990, 31,5153. 4.1 g of the compound of the above step are suspended in 50 ml of6N HCl and the mixture is refluxed for 5 hours. It is concentrated inorder to obtain a first crop of the pure expected compound. Onevaporating to dryness, a second crop of 1.4 g of the expected compoundcontaminated with NH₄ Cl is obtained. A sample is treated with (Boc)₂ Oand its optical rotation is measured.

α_(D) ²⁵ =-17° (c=1; MeOH)

literature α_(D) ²⁵ =-16° (c=1; MeOH)

Preparation 2.21

3-Amino-3-phenylpropionic acid trifluoroacetate, (S) isomer.

The preparation is identical to the one described in 2.19, but usingD(+)-menthol.

Preparation 3.1

3-Phenyl-3-(2,4,6-trichlorobenzenesulphonamido-)propionic acid.

1.15 g of 3-amino-3-phenylpropionic acid are dissolved in 25 ml ofdioxane and 7 ml of 1N sodium hydroxide and 1.95 g of2,4,6-trichlorobenzenesulphonyl chloride in 5 ml of dioxane aregradually added, while. maintaining the pH at 10.5-11 by addition of 1Nsodium hydroxide. After stirring for 2 hours at RT, the mixture isdiluted with 100 ml of water, extracted twice with EtOAc and acidifiedto pH=2 by addition of 6N HCl. The solid formed is drained, washed withwater and dried at 40° C. 2.09 g of the expected product are obtained,m.p.=218-219° C.

NMR (DMSO+TFA) : 2.60-2.95 : mt : 2H; 4.75 : q : 1H; 7.10-7.30 : m : 5H;7.60 : s : 2H; 8.85 : d : 1H.

Preparation 3.2

3-(Naphth-2-ylsulphonamido)-3-phenylpropionic acid.

4.13 g of 3-amino-3-phenylpropionic acid are dissolved in 100 ml ofdioxane and 25 ml of IN sodium hydroxide and 5.6 g of2-naphthalenesulphonyl chloride are added portionwise, while maintainingthe pH at 10.5-11 by addition of 1N sodium hydroxide. After stirring for2 hours at RT, the mixture is diluted with 400 ml of water and 2N HCl isadded in order to obtain pH=2. This mixture is extracted with EtOAc andthe extracts are washed with a KHSO₄ /K₂ SO₄ buffer, dried over Na₂ SO₄and evaporated to dryness. The residue is triturated from heptane andthe product is isolated by decanting and is then dried under vacuum.7.33 g of the expected product are obtained, m.p.=126-129° C.

NMR (DMSO+TFA) : 2.55-2.70 : mt : 2H; 4.70 : t : 1H; 6.85-8.15 : m :12H.

Preparation 3.3

2-Hydroxy-3-(naphth-2-ylsulphonamido)-3-phenylpropionic acid.

This compound is prepared according to Bull. Soc. Chim., France, 1940,593-603.

Working according to the procedures described in Preparations 3.1 and3.2, starting with the compounds obtained in Preparation 2 and2-naphthalenesulphonyl chloride, the acids described in Table 4 beloware obtained.

                  TABLE 4                                                         ______________________________________                                         ##STR52##                                                                    Preparations R.sub.2    NMR         m.p. ° C.                          ______________________________________                                        3.4                                                                                    ##STR53##      (DMSO)  2.80: d: 2H; 4.90:  q: 1H; 7.15-8.20:  m:                             14H; 8.60:  d: 1H                                                                         145-150                                   3.5                                                                                    ##STR54##      (DMSO + TFA)  2.00-2.20: mt:  2H; 4.45: t: 1H;                                7.10-8.25:  m: 11H                                                                        223-228                                   3.6                                                                                    ##STR55##      (DMSO)  2.50-2.75: mt:  2H; 4.60-4.80:  mt: 3H                                6.65-8.15: m:  16H 8.40: bs: 1H                                                           253-255                                   3.7                                                                                    ##STR56##      RMN(DMSO)  2.40-2.65: mt:  2H 4.65: s: 2H;  4.75:                             dq: 1H;  6.40-8.05: m:  16H 8.40: bs:                 ______________________________________                                                                1H                                                

The acids described above are converted by the action ofN-hydroxysuccinimide in the presence of DCC in DMF. The compoundsobtained are described in Table 5 below.

                  TABLE 5                                                         ______________________________________                                         ##STR57##                                                                    Preparations                                                                          R.sub.2         NMR(DMSO)   m.p.(° C.)                         ______________________________________                                        3.8                                                                                    ##STR58##      s.75: s: 4H; 3.20:  mt: 2H; 4.90: dq:  1H;                                    7.20-8.10:  m: 14H; 8.70:  d: 1H                                                          165                                       3.9                                                                                    ##STR59##                  192                                       3.10                                                                                   ##STR60##                  187                                       3.11                                                                                   ##STR61##      2.80: s: 4H; 3.15:  mt: 2H; 4.65: s:  2H; 4.75:                               dq: 1H;  6.45-7.95: mt:  4H; 7.25-8.20:  mt: 12H;                             8.60:  d: 1H                                                                              110                                       ______________________________________                                    

Preparation 3.12

3-(2-Hydroxyphenyl)-3-(naphth-2-ylsulphonamido)propionic acid.

1.1 g of 3-amino-3-(2-hydroxyphenyl)propionic acid hydrochloride,prepared according to J. Agric. Food Chem., 1977, 25, 965, are dissolvedin 25 ml of dioxane and 10 ml of 1N NaOH are added; 1.12 g of2-naphthalenesulphonyl chloride are gradually added while maintainingthe pH at 11.5-12 by addition of 1N NaOH. After stirring for 2 hours atRT, the reaction medium is diluted in water, washed with EtOAc (twice)and then acidified to pH=1.5-2 by addition of 6N HCl. The whiteprecipitate formed is drained, washed with water and dried in order toobtain 0.82 g of the expected product, m.p.=190-200° C.

NMR (DMSO) 2.50-3.20 : mt : 2H; 4.80 : mt : 1H; 6.95-8.30 : m : 11H;8.75 : s : 1H.

Preparation 3.13

3-Phenyl-3-(quinol-2-ylsulphonamido)propionic acid.

A) 2-Quinolinesulphonyl chloride.

3.2 g of 2-mercaptoquinoline are suspended in 40 ml of water containing0.156 g of iron trichloride. The mixture is cooled in an ice bath andchlorine is then bubbled through for 1 hour at 4° C. The mixture isevaporated and the residue is then taken up in the minimum amount ofwater and the product is drained and dried in order to obtain 2.30 g ofthe expected compound in dry form.

B) 3-Phenyl-3-(quinol-2-ylsulphonamido)propionic acid.

1.65 g of the compound prepared in the above step are dissolved in 40 mlof dioxane and 20 ml of 1N NaOH are added, along with portionwiseaddition of 2.27 g of 3-amino-3-phenylpropionic acid, while maintainingthe pH at 10. The mixture is left stirring for 18 hours at RT and isthen evaporated to dryness. The residue is taken up in a DCM/H₂ Omixture, the phases are separated after settling has taken place and theorganic phase is extracted with water. Acidification to pH=1 is carriedout by addition of HCl and the product is then drained and dried overMgSO₄ in order to obtain 1.9 g of the expected compound.

NMR (DMSO+TFA) : 2.4-2.9 : m : 2H; 4.9 : mt : 1H; 6.9-7.1 : mt : 3H; 7.2: d : 2H; 7.7-8.1 : m : 5H; 8.4 : d : 1H; 8.8 d : 1H.

Preparation 3.14

3-Phenyl-3-(quinol-8-ylsulphonamido)propionic acid.

This compound is prepared according to the procedure described above,starting with 8-quinolinesulphonyl chloride.

Preparation 3.15

3-(3-Isopropyloxyphenyl)-3-(naphth-2-ylsulphonamido)propionic acid.

1.3 g of the compound of Preparation 2.7 are suspended in 20 ml ofdioxane and treated with 2N NaOH and then with 1N NaOH in order to bringthe mixture to pH=12. 1.13 g of 2-naphthalenesulphonyl chloride areadded portionwise, while maintaining the pH at 10.5-11.5 by addition of1N NaOH. The mixture is left stirring for 2 hours at RT and is thendiluted with water, washed with EtOAc and then brought to pH=2 byaddition of 2N HCl. This mixture is extracted with EtOAc and then washedwith a KHSO₄ /K₂ SO₄ buffer and with saturated NaCl solution. Theresulting solution is dried over Na₂ SO₄ and concentrated in order toobtain 900 mg of the expected compound, m.p.=126° C.

NMR (DMSO) : 0.95 : d : 6H; 2.40-2.55 : mt : 2H; 4.10 : tq : 1H; 4.60 dq:H; 6.20-8.05 : mt : 11H; 8.30 : d : 1H; 12.20 : bs : 1H.

Following the procedure described in the above preparation, and usingthe compounds of Preparations 2.8 to 2.15 as starting materials, thecompounds described in Table 6 below are prepared.

                  TABLE 6                                                         ______________________________________                                         ##STR62##                                                                    Preparation                                                                           R.sub.2         NMR         m.p. ° C.                          ______________________________________                                        3.16                                                                                   ##STR63##      (DMSO)  2.35-2.55: mt:  2H; 3.35: s: 3H;  4.60:                               dq: 1H;  6.20-6.85: mt:  4H; 7.40-8.0:  mt: 7H;                               8.30: d:  1H; 12.20: bs: 1H                                                               154                                       3.17                                                                                   ##STR64##      (DMSO)  2.35-2.60: mt:  2H; 3.35: s: 3H;  4.55:                               dq: 1H;  6.35: d: 2H; 6.90:  d: 2H;  7.40-7.95:                               mt:  7H; 8.25: d: 1H;  12.20: bs:                                                         139                                       3.18                                                                                   ##STR65##      (DMSO)  2.40-2.70: mt:  2H; 4.60: mt: 1H;                                     6.70-7.10: m:  4H; 7.40-8.20:  m: 7H; ; 8.45: bd:                              1H; 12.30: bs: 1H                                                                        133                                       3.19                                                                                   ##STR66##      (DMSO)  2.40-2.70: mt:  2H; 4.70: dq: 1H;                                     7.00-8.00: m:  11H; 8.45: d: 1H;  12.20: bs:                                              164                                       3.20                                                                                   ##STR67##      (DMSO)  2.80: bd: 2H;  5.60: mt: 1H;  7.10-8.10:                              m:  14H; 8.65: d: 1H;  12.30: bs:                                                         189                                       3.21                                                                                   ##STR68##      (DMSO)  2.45-2.65: mt:  2H; 4.70: dq: 1H                                      6.60-7.05: m:  4H; 7.50-8.15:  m: 7H; 8.50: d:                                1H; 12.30: bs: 1H                                                                         167                                       3.22                                                                                   ##STR69##      (DMSO + TFA)  0.80-1.60: m:  11H; 1.85-2.30:  mt:                             2H; 3.40: dq:  1H; 7.50-8.30:  m:                                                         198                                       3.23                                                                                   ##STR70##      (DMSO) 1.85: s:  3H; 2.45-2.65:  mt: 2H; 4.65:                                dq:  1H; 6.60-6.90:  m: 4H;  7.50-8.15: m:  7H;                               8.40: bd: 1H;  12.20: bs: 1H                                                              102                                       ______________________________________                                    

The acids described above (Preparations 3.15 to 3.23) are converted bythe action of N-hydroxysuccinimide in the presence of DCC in dioxane.The compounds obtained are described in Table 7 below.

                  TABLE 7                                                         ______________________________________                                         ##STR71##                                                                    Preparations                                                                          R.sub.2         NMR         m.p. ° C.                          ______________________________________                                        3.24                                                                                   ##STR72##      --           97                                       3.25                                                                                   ##STR73##      --          142                                       3.26                                                                                   ##STR74##      --          135                                       3.27                                                                                   ##STR75##      --          163                                       3.28                                                                                   ##STR76##      --          142                                       3.29                                                                                   ##STR77##      (DMSO + TFA)  2.70: s: 4H; 3.25:  d: 2H; 5.60: t:                              1H; 7.05-7.95:  m: 14H                                                                   192                                       3.30                                                                                   ##STR78##      (DMSO)  2.70: s: 4H;  2.95-3.10: mt:  2H; 4.75:                               dq: 1H;  6.50-6.90: m:  4H; 7.45-8.10:  m: 7H;                                8.60:  d: 1H                                                                              174                                       3.31                                                                                   ##STR79##      (DMSO + TFA)  0.60-1.60: m:  11H; 2.40-2.80:  mt:                             2H; 2.70: s:  4H; 3.35: mt: 1H;  7.50-8.30: m:                                            135                                       3.32                                                                                   ##STR80##      --          142                                       ______________________________________                                    

Preparation 3.33

3-(3,4-Dichlorobenzenesulphonamido)-3-phenylpropionic acid.

A) 3,4-Dichlorophenylbenzenesulphonyl chloride.

5.4 g of (3,4-dichloro)thiophenol are suspended in 60 ml of water; 0.234g of FeCl₃ is added and chlorine is bubbled through for one hour at atemperature below 10° C. The mixture is evaporated, drained, washed withwater and then dried by azeotropic entrainment in order to obtain 6.7 gof the expected compound.

B) 3-(3,4-Dichlorobenzenesulphonamido)-3-phenylpropionic acid.

The process is then carried out according to the procedure ofPreparation 3.1 in order to obtain the expected compound.

NMR (DMSO) : 2.5-2.9 : m : 2H; 4.6-4.8 : q : 1H; 7.1 : s : 5H; 7.5-7.7 :mt : 3H; 8.7 : d : 1H; 12.4 : s : 1H.

Preparation 3.34

3-(5,6,7,8-Tetrahydronaphth-2-ylsulphonamido)-3-phenylpropionic acid.

A) Sodium 5,6,7,8-tetrahydronaphthalene-2-sulphonate.

3.9 g of 1,2,3,4-tetrahydronaphthalene are dissolved-in 10 ml ofanhydrous CCl₄ at 0° C.; 4.3 g of dioxane in 10 ml of CCl₄ are added,followed by 6.1 ml of sulphuric anhydride. The mixture is left stirringfor 18 hours at RT and then for 3 hours at 80° C. A water/ice mixture isadded and the resulting mixture is extracted with ether. The aqueousphase is adjusted to pH=6.5 by addition of 5N NaOH. The product isdrained and dried over Na₂ SO₄ and then by azeotropic entrainment. 10.6g are obtained.

B) 5,6,7,8-Tetrahydronaphthalene-2-sulphonyl chloride.

2 g of the sulphonate obtained in the above step are mixed with 5 g ofphosphorus pentachloride and the mixture is refluxed for 6 hours. It isevaporated, poured onto a water/ice mixture and then extracted with DCM,dried over Na₂ SO₄ and evaporated in order to obtain 1.5 g of theexpected compound.

C) 3-(5,6,7,8-Tetrahydronaphth-2-ylsulphonamido)-3-phenylpropionic acid.

The process is then performed according to the procedure of Preparation3.1 in order to obtain the expected product.

NMR (DMSO) : 1.6-1.8 : m : 4H; 2.5-2.8 : m : 6H; 4.5-4.7 : dd : 1H;7-7.2 : m : 7H; 7.3 : d : 1H; 8.1 : d : 1H; 12.3 : s : 1H.

Preparation 3.35

1-(Naphth-2-ylsulphonamido)indane-2-carboxylic acid.

A) Methyl (1-oxo)indan-2-carboxylate.

The reaction is carried out according to J. Med. Chem., 1970, 650. Asuspension of 6.75 g of sodium hydride at 80% in oil and 45 g of methylcarbonate in 120 ml of benzene are mixed together. 11.9 g of 1-indanonedissolved in 100 ml of benzene are added, at 60° C., over 1 and a halfhours and the mixture is then refluxed. After 1 hour, the benzene isdistilled off. Xylene is added and the mixture is again refluxed. After1 hour, it is cooled, 30 ml of AcOH are added and the resulting mixtureis poured onto 200 ml of a water/ice mixture containing 30 ml of 1N HCl.The insoluble material is removed by filtration and the filtrate isextracted with Et₂ O. The extracts are washed with water, with saturatedNaHCO₃ solution, with water, with saturated NaCl solution and then driedover Na₂ SO₄. The product is chromatographed on silica, eluting withEtOAc/hexane (1.3; v/v) in order to obtain 3.05 g of the expectedcompound.

NMR (DMSO) : 3.25-3.50 : mt : 2H; 3.70 : ds : 3H; 4.85-4.90 : mt : 1H;7.40-7.80 : m : 4H. Multiple signals since the product is partly inenolic form.

Comment using THF as solvent instead of benzene and xylene, a betteryield is obtained: 67% instead of 30%.

B) Methyl 1-hydroxyiminoindan-2-carboxylate.

The reaction is carried out according to J. Heterocycl. Chem., 1974, 11,982. 3.04 g of the compound of the above step in 9 ml of MeOH are addedover 10 minutes to a mixture of 2.21 g of sodium acetate and 3.1 g ofhydroxylamine hydrochloride in 3 ml of water. The mixture is thenrefluxed for 1 hour 30 minutes, after which it is cooled and extractedwith EtOAc. The extracts are washed with water, with 3/4 saturatedNaHCO₃ solution, with water and with saturated NaCl solution. Afterevaporating off the EtOAc, 3.28 g of the expected compound are obtainedin solid form.

C) Methyl 1-aminoindan-2-carboxylate hydrochloride.

3.27 g of the compound of the above step are dissolved in 80 ml of EtOH,1.2 g of 10% Pd/C are added, followed by 20 ml of 1M hydrochloricethanol and the mixture is left stirring under a pressure of hydrogen (5bar) at RT overnight. The catalyst is filtered off, the filtrate isconcentrated under vacuum and the residue is then taken up in EtOAc andwater. The water is separated out after settling of the phases has takenplace and is then brought to pH=8.5 by addition of 2N NaOH. This phaseis extracted with EtOAc, washed with water, with saturated NaClsolution, dried over Na₂ SO₄ and evaporated. The residue is taken up in20 ml of MeOH and 6 ml of 2.5N HCl. The mixture is concentrated undervacuum and the residue is then triturated from Et₂ O. 2.71 g of theexpected compound are obtained in the form of a powder.

D) Methyl 1-(2-naphthalenesulphonamido)indan-2-carboxylate.

0.91 g of the above compound is placed in 10 ml of chloroform and 0.95 gof 2-naphthalenesulphonyl chloride is then added portionwise, followedby portionwise addition of 0.68 ml of DIPEA in order to maintain themixture at pH=7-8. After 3 hours, the reaction medium is extracted withEtOAc and then washed with 0.05N NaOH, 0.25N HCl, with water and withsaturated NaCl solution. 0.99 g of the expected compound is obtained inthe form of a pink-white solid.

NMR (DMSO+TFA) : 2.80-3.30 : mt : 2H; 3.40 : s : 3H; 3.50 : q : 1H; 5.10: d : 1H; 6.40-8.50 : mt : 11H.

E) 1-(Naphth-2-ylsulphonamido)indan-2-carboxylic acid.

0.98 g of the compound of the above step is placed in 10 ml of MeOH, 1ml of 8.36N KOH is added, the mixture is heated at 50° C. for 3 hoursand a further 0.25 ml of 8.36N KOH is then added. After 2 hours, themedium is diluted by addition of water and EtOAc and the pH is broughtto 2.5 by addition of 1N HCl. The organic phase is separated out aftersettling of the phases has taken place, washed with H₂ O, with saturatedNaCl solution and then dried over Na₂ SO₄ and evaporated. 0.80 g of theexpected compound is obtained in the form of a foam.

Preparation 3.36

[2-(2-Naphthalenesulphonyl)-1,2,3,4-tetrahydro-1-isoquinolyl]aceticacid.

1.91 g of (1,2,3,4-tetrahydro-1-isoquinolyl)acetic acid (Preparation2.18) are suspended in 25 ml of dioxane, 10 ml of 1N NaOH are added,followed by portionwise addition of 2.3 g of 2-naphthalenesulphonylchloride, and the mixture is maintained at pH=10.5-12 by addition of 1NNaOH. The mixture is kept stirring at a constant pH for 2 hours at RT.The reaction medium is diluted with 100 ml of water and is then washedtwice with EtOAc and brought to pH=1.2 by addition of 6N HCl in thepresence of EtOAc. This mixture is extracted with EtOAc, washed withsaturated NaCl solution, dried over Na₂ SO₄ and evaporated to dryness.The expected product crystallizes from heptane, and is drained, washedwith heptane and then dried to give 3.14 g, m.p.=132° C.

NMR (DMSO+TFA) : 2.40-2.80 : m : 4H; 3.40-3.90 : m : 2H; 5.45 : t : 1H;6.90-7.20 : m : 4H; 7.50-8.40 : m : 7H.

Preparation 3.37

The 2,5-dioxo-1-pyrrolidinyl ester of the above acid is prepared by theaction of NSuOH in the presence of DCC in dioxane.

NMR (DMSO+TFA) : 2.50-2.70 : m : 2H; 2.75 : s : 4H; 3.10-3.35 : mt : 2H;3.50-3.80 : mt : 2H; 5.50 : t : 1H; 6.80-8.40 : m : 11H.

Preparation 3.38

3-(2-Naphthalenesulphonamido) -3-phenylpropionic acid, (R) isomer.

2 g of the compound of step 2.19 are placed in 30 ml of dioxane in thepresence of 7.2 ml of 2N NaOH; 1.6 g of 2-naphthalenesulphonyl chlorideare added portionwise while maintaining the mixture at pH=10.5-11.5 byaddition of 1N NaOH. After stirring for 2 hours at RT, the mixture isdiluted by addition of 100 ml of water and is then washed with EtOAc(several times). The aqueous phase is diluted with EtOAc and thentreated with 2N HCl until the pH=2.2. The resulting mixture is extractedwith EtOAc, washed with saturated NaCl solution, dried over Na₂ SO₄ andevaporated to dryness. The expected compound crystallizes from heptane,and is drained, washed with heptane and then dried. 2.2 g are obtained,m.p.=123-126° C.

α_(D) ²⁵ =+67.9 (c=1; MeOH)

Preparation 3.39

2,5-Dioxo-1-pyrrolidinyl3-(2-naphthalenesulphonamido)-3-phenylpropionate, (R) isomer.

A solution of 2 g of the above compound and 657 mg ofN-hydroxysuccinimide in 35 ml of dioxane is prepared and 1.24 g of DCCIin 10 ml of dioxane are gradually added. After stirring for 5 hours atRT, the DCU is drained and washed with acetone and the filtrate is thenevaporated to dryness. The residue is taken up in isopropanol. Theproduct which crystallizes is drained, washed with Et₂ O and then dried.2.18 g of the expected compound are obtained.

Preparations 3.40 and 3.41

The process is performed as in the 2 preparations described above,starting with the compound of Preparation 2.21 in order to obtain3-(2-naphthalenesulphonamido)-3-phenylpropionic acid, (S) isomer and its2,5-dioxo-1-pyrrolidinyl ester.

Preparation 3.42

3-(4-Chlorophenyl)-3-(2-naphthalenesulphonamido)propionic acid, (R)isomer.

A) 3-(4-Chlorophenyl)-3-(phenylacetamido)propionic acid.

9.6 g of 3-amino-3-(4-chlorophenyl)propionic acid are dissolved in 200ml of dioxane and about 50 ml of 1N NaOH in order to reach pH=10.5-11.5.The medium is cooled to +5° C. and 6.34 ml of phenylacetyl chloride arethen gradually added, while maintaining the mixture at pH=10.5-11.5 andat a temperature of between +5° C. and +10° C. Stirring is continued for1 and a half hours and the medium is then concentrated to one-halfbefore diluting with 500 ml of water. The resulting mixture is washedtwice with EtOAc and then acidified to pH=1-2 by addition of 6N HCl. Thesolid is drained, washed with water and dried to give 14 g of theexpected compound.

B) 3-Amino-3-(4-chlorophenyl)propionic acid, (R) isomer.

This step is carried out according to Syn. Letters, 1993, 339. 16.8 g ofthe compound of the above step are suspended in 300 ml of water and 1NNH₄ OH is added until a pH=7.5 is obtained. 0.6 ml of Sigmapenicillamidase® is added and the mixture is left stirring for 72 hoursat 40° C. The expected compound precipitates. It is drained, washed withwater and with acetone and then dried at 40° C. 2.85 g are obtained.

α_(D) ²⁵ =-5° (c=1, 1N HCl)

C) 3-(4-Chlorophenyl)-3-(2-naphthalenesulphonamido)propionic acid, (R)isomer.

1 g of the compound obtained in the above step is dissolved in 15 ml ofdioxane and 5 ml of 1N NaOH in order to reach pH=10.5-11.5. 1.15 g of2-naphthalenesulphonyl chloride are gradually added while keeping the pHconstant. After stirring for 2 hours at RT, the medium is diluted byaddition of an equal volume of water and is then washed twice withEtOAc. The medium is acidified to pH=1.3 by addition of 6N HCl. Thismixture is extracted with EtOAc and the extracts are washed withsaturated NaCl solution (several times) The resulting solution is driedover Na₂ SO₄ and evaporated to dryness, and the expected product thencrystallizes from heptane. 1.31 g are obtained.

α_(D) ²⁵ =+92° (c=1; MeOH)

Preparation 3.43

The 2,5-dioxo-1-pyrrolidinyl ester of the above acid is preparedaccording to the usual technique.

Preparation 3.44

4-Phenyl-3-(2-naphthalenesulphonamido)butanoic acid, (S) isomer.

1.08 g of the compound of Preparation 2.20 are dissolved in 30 ml ofdioxane and 10 ml of water. 10N NaOH is added in order to reach pH=14and 1.13 g of 2-naphthalenesulphonyl chloride are then addedportionwise, while maintaining the mixture at pH=10.5-11.5. Stirring iscontinued for 2 hours and the mixture is then diluted with water andwashed with EtOAc. It is again diluted with EtOAc and acidified to pH=2by addition of 2N HCl and extracted with EtOAc; the extracts are washedwith saturated NaCl solution (several times) and then dried andevaporated to dryness. On addition of heptane, the expected productcrystallizes. 1.25 g are obtained.

α_(D) ²⁵ =-26.8 (c=1; MeOH)

Preparation 3.45

The 2,5-dioxo-1-pyrrolidinyl ester of the above acid is prepared usingthe usual techniques.

Preparation 3.46

(2S,4R) (4-(Benzyloxy)-1-(2-naphthalenesulphonyl)-2-pyrrolidinyl)aceticacid.

A) 4-(Hydroxy)-1-(2-naphthalenesulphonyl)pyrrolidine-2-carboxylic acid.

2.62 g of (2S,4R)-4-hydroxyproline are dissolved in 15 ml of watercontaining 5.9 g of Na₂ CO₃ and 5.21 g of 2-naphthalenesulphonylchloride are added. After vigorous stirring for 18 hours at RT, themixture is drained and the filtrate is then acidified to pH=1. Theresulting mixture is again drained, washed with water and dried in orderto obtain 4.1 g of the expected compound.

B) 4-(Benzyloxy)-1-(2-naphthalenesulphonyl)2-pyrrolidinylacetic acid.

1 g of the compound of the above step is dissolved in 20 ml of DMF at 0°C. under nitrogen; 0.204 g of sodium hydride at 80% in oil is added andthe mixture is then left stirring for 1 hour at 0° C. and for 30 minutesat RT. The mixture is cooled to 0° C. and 10 mg of 16-crown-6 crownether and 0.921 ml of benzyl bromide are then added. The mixture is leftstirring for 1 hour at 0° C and then for 18 hours at 50° C. and 10 ml of1N NaOH are added. After stirring for 18 hours at RT, the mixture isdiluted with water and then extracted with ether and acidified to pH=2by addition of 1N HCl. The resulting mixture is extracted with EtOAc andthe extracts are dried over Na₂ SO₄ and evaporated in order to obtain0.8 g of the expected compound.

C)(2S,4R)-4-Benzyloxy-2-hydroxymethyl-1-(2-naphthalenesulphonyl)pyrrolidine.

2.8 g of the compound of the above step are dissolved in 20 ml of THF.1.12 ml of Et₃ N and 0.968 ml of ethyl chloroformate are added over 30minutes at RT, followed by addition of 0.762 g of sodium borohydride in5 ml of water. After stirring for 20 hours at RT, the mixture isevaporated and the residue is taken up in 20 ml of water and acidifiedto pH=3 by addition of 1N HCl. The product is drained, washed withwater, dried and then precipitated from an EtOAc/hexane mixture in orderto obtain 1.9 g of the expected compound.

D)(2S,4R)-4-Benzyloxy-2-mesyloxymethyl-1-(2-naphthalenesulphonyl)pyrrolidine

3 g of the compound of the above step are dissolved in 100 ml of DCM at0° C. and 1.28 ml of Et₃ N and then 0.72 ml of mesyl chloride are added.After stirring for 30 minutes at 0° C., a further 1.28 ml of Et₃ N andthen 0.72 ml of mesyl chloride are added, after which the mixture isleft stirring for 30 minutes at RT. It is washed, while cold, with KHSO₄/K₂ SO₄ and the insoluble material is then filtered off and the filtrateis dried and evaporated. The residue is taken up in Et₂ O and usedwithout further purification in the following step.

E) (2S,4R)4-Benzyloxy-2-cyanomethyl-2-(2-naphthalenesulphonyl)pyrrolidine.

The product of the above step is dissolved in

50 ml of DMSO, 2 g of 10-crown-6 crown ether are added and the mixtureis cooled on an ice bath. 5 g of potassium cyanide are then added andthe mixture is heated for 4 hours at 50° C. The medium is diluted with250 ml of EtOAc and is then washed with water, dried and evaporated. Theresidue is taken up in ether. The expected compound crystallizes and 1.7g are obtained.

F) Ethyl (2S,4R)(4-(benzyloxy)-1-(2-naphthalenesulphonyl)-2-pyrrolidinyl)acetate.

1.6 g of the compound of the above step are dissolved in 50 ml ofHCl-saturated ethanol at 0° C. The mixture is left stirring for 48 hoursat +4° C. and is then evaporated. The residue is taken up in EtOH andevaporated (twice) and then taken up in Et₂ O and evaporated (twice). 20ml of boiling water are added and a few ml of dioxane/EtOH mixture arethen added to homogenize it. The solution is heated for 15 minutes at100° C. This mixture is evaporated and the residue is taken up indioxane and then neutralized to pH 6 by addition of 1N NaOH. Thesolution is used without further purification in the following step.

G) (2S,4R)(4-Benzyloxy)-1-(2-naphthalenesulphonyl)-2-pyrrolidinyl)acetic acid.

2.4 ml of 5N NaOH are added to the solution of the above step and themixture is heated at 50° C. for 30 minutes. The solvent is evaporatedoff and the residue is then acidified to pH=3 by addition ofconcentrated HCl. The product is drained, washed with water and dried inorder to obtain 1.44 g of the expected compound.

Preparation 4.1

A)N-(2-(4-(3,4-Dihydroimidazol-2-yl)phenyl)-1-((1-pyrrolidinyl)carbonyl)ethyl)-3-(3,4-dichlorophenyl)-3-(N-Boc)aminopropionamide.

1.09 g of1-[2-amino-3-(4-(3,4-dihydroimidazol-2-yl)phenylpropionyl]pyrrolidinedihydrochloride are dissolved in 15 ml of DMF, 350 μl of Et₃ N are addedand the mixture is stirred for a few minutes before adding 835 mg of3-((N-Boc)amino)-3-(3,4-dichlorophenyl)propionic acid (obtained inPreparation 2.6) and 515 mg of DCC.

After stirring for 3 hours, the mixture is evaporated to dryness and theresidue is then taken up in methanol. The DCU is drained and thefiltrate is concentrated to one-half and diluted with acetone. Thismixture is filtered and then evaporated to dryness. The residue ischromatographed, eluting with a chloroform/methanol mixture (from100/2.5 to 100/20; v/v).

B)N-(2-(4-(3,4-Dihydroimidazol-2-yl)phenyl)-1-((1-pyrrolidinyl)carbonyl)ethyl)-3-(3,4-dichlorophenyl)-3-aminopropionamide.

The crude product obtained in the above step is placed in 20 ml of asolution of 4N HCl in dioxane and a sufficient amount of MeOH to allowcomplete dissolution. After 1 hour at RT, the mixture is evaporated todryness and the residue is triturated twice from ether and then drained,washed with ether and dried under vacuum in the presence of KOH. 225 mgof the expected compound are obtained.

Preparation 4.2

N-[2-(4-Cyanophenyl)-1-((1-pyrrolidinyl)carbonyl)ethyl]-3-phenyl-3-aminopropionamidetrifluoroacetate, (R,R) isomer.

A)N-[2-(4-Cyanophenyl)-1-((1-pyrrolidinyl)carbonyl)ethyl]-3-phenyl-3-(N-Boc)aminopropionamide,(R,R) isomer.

The 2,5-dioxo-1-pyrrolidinyl ester of the acid obtained in Preparation2.19, step B is prepared. 800 mg of this compound are added to a mixturecontaining 790 mg of the compound of Preparation 1.13 in 15 ml ofacetonitrile and 380 μl of DIPEA. The mixture is left stirring for a fewhours and is then left overnight at RT. The medium is evaporated todryness and the residue is taken up in KHSO₄ /K₂ SO₄ and then extractedwith EtOAc. The organic phase is washed with saturated NaCl solution andthen dried over Na₂ SO₄ and evaporated to dryness. 1.1 g of the expectedcompound are obtained, this compound crystallizing from heptane.

B)N-[2-(4-Cyanophenyl)-1-((1-pyrrolidinyl)carbonyl)ethyl]-3-phenyl-3-aminopropionamidetrifluoroacetate, (R,R) isomer.

1 g of the compound of the above step is dissolved in 6 ml of DCM, 6 mlof TFA are then added and the mixture is left stirring for 35 minutes atRT. The reaction medium is evaporated to dryness, the residue isredissolved in isopropanol and then evaporated, the residue istriturated from Et₂ O and the product is drained and dried in order toobtain 0.88 g of the expected compound.

EXAMPLE 1 ##STR81##

210 mg of the compound of Preparation 4.1 are dissolved in 10 ml ofdioxane and 5 ml of water, and IN sodium hydroxide is added in order toreach pH=9.5. 84 mg of 2-naphthalenesulphonyl chloride are added 3times, while maintaining the mixture at pH=9-9.5 by addition of 0.5Nsodium hydroxide. After leaving overnight at RT, the medium is dilutedwith water and acidified to pH=2.5 by addition of 1N HCl. The solidformed is chromatographed on Sephadex® LH 20, eluting with methanol. Thefraction containing the expected product is concentrated, treated with asolution of 4N HCl in dioxane and then evaporated to dryness; theresidue is taken up in ether, drained and dried. 41 mg of the expectedcompound are obtained.

MH⁺ : 692 : dichloro isotopic profile

NMR (DMSO+TFA) 1.40-1.70 : m : 4H; 2.50-3.40 : m : 8H; 4.00 : ds : 4H;4.45-4.70 : m : 2H; 6.60-8.00 : m : 14H

EXAMPLE 2 ##STR82##

715 mg of the compound of Preparation 1.1, 15 ml of CH₃ CN, 250 μl oftriethylamine, 710 mg of the compound of Preparation 3.2 and 450 mg ofDCC are mixed together and left stirring for 5 hours at RT. The mixtureis evaporated to dryness, the residue is taken up in acetone, the DCU isdrained and the filtrate is then evaporated to dryness again. Theresidue is triturated from ether and the product is then isolated bydecanting, several times. It is dried under vacuum in the presence of P₂O₅ in order to obtain 610 mg of the expected compound, m.p.=195-200° C.

NMR (DMSO+TFA) 1.40-1.70 : m : 4H; 2.30-3.40 : m : 8H; 4.40-4.60 : m :and 4.60-4.70 : m : 2H; 6.75-8.15 m : 16H

B)

600 mg of the product obtained in the above step are dissolved in 20 mlof a solution of ethanol saturated with hydrochloric acid, at 0° C.After leaving overnight in a refrigerator, the mixture is evaporated todryness and the residue is then dried under vacuum in the presence ofKOH. The product obtained is diluted in 25 ml of anhydrous ethanol and134 μl of ethylenediamine in 2 ml of anhydrous EtOH are then addedseveral times. After leaving overnight at RT, the mixture is evaporatedto dryness and the residue is then chromatographed on silica, elutingwith a chloroform/methanol mixture (85/15; v/v). The fractionscontaining the expected compound are combined, evaporated and then takenup in dilute hydrochloric ether. After draining and drying under vacuumin the presence of KOH, 180 mg of the expected product are obtained.

MH⁺ : 624

NMR (DMSO+TFA) 1.40-1.80 : m : 4H; 2.55-3.45 : m : 8H; 4.00 : s : 4H;4.55 : bt : and 4.70 : bt : 2H; 6.70-8.20 : m : 16H

EXAMPLE 3 ##STR83## A)

800 mg of the compound obtained in Example 2, step A are dissolved in 20ml of a saturated solution of HCl in anhydrous EtOH. After leavingovernight in a refrigerator, the mixture is evaporated to dryness andthe residue is then dried under vacuum in the presence of KOH. Theresidue is dissolved in 30 ml of anhydrous EtOH and 175 μl of Et₃ N areadded, followed by 350 mg of N-Boc-(4-aminomethyl)benzylamine. After 48hours at RT, the mixture is evaporated to dryness and the residue isthen chromatographed on Sephadex® LH 20, eluting with MeOH. 2 fractionscontaining two different compounds are obtained. For one (fraction 1,570 mg), the amidine is monosubstituted with a group R₈=-4((N-Boc)aminomethyl)benzyl and R₆ and R₇ =H; for the other (fraction2, 150 mg), the amidine is disubstituted with R₆=R=-4(N-(Boc)aminomethyl)benzyl and R₇ =H.

B)

125 mg of the compound of fraction 2 obtained in the above step aresuspended in 1 ml of DCM, 1 ml of TFA is added and the mixture is leftstirring for 40 minutes at RT. After evaporating the medium to dryness,the residue is dissolved in isopropanol and is then evaporated todryness again; the residue is triturated from Et₂ O, drained, washedwith Et₂ O and then dried. 110 mg of the expected compound are obtained.

MH⁺ : 836

NMR (DMSO+TFA) 1.40-1.80 : m : 4H; 2.40-3.30 : m : 8H; 3.90-4.80 : m :10H; 6.80-8.45 : m : 24H

EXAMPLE 4 ##STR84##

535 mg of the compound of fraction 1 obtained in Example 3, step A aresuspended in 8 ml of DCM, 8 ml of TFA are added and the mixture is leftstirring for 40 minutes at RT. After evaporating the medium to dryness,the residue is triturated from ether and then dried under vacuum in thepresence of potassium hydroxide. 520 mg of the expected compound areobtained.

MH⁺ : 717

NMR (DMSO+TFA) ; 1.30-1.70 : m : 4H; 2.20-3.30 : m : 8H; 3.90 : s : 2H;4.40-4.70 : m : 4H; 6.80-8.00 : m : 20H

EXAMPLE 5 ##STR85##

960 mg of the compound of Preparation 3.8 in 20 ml of DMF are mixed with852 mg of the compound of Preparation 1.2 and 280 Al of Et₃ N and themixture is adjusted to pH=7-7.5 by addition of Et₃ N. After stirringovernight, the mixture is evaporated to dryness and the residue is thentaken up in methanol and chromatographed on Sephadex® LH 20, elutingwith methanol. The fractions containing the product are combined,evaporated and then taken up in 20 ml of butanol, 10 ml of 1N HCl and 10ml of water. After stirring and separation of the phases by settling,the organic phase is evaporated. The residue is chromatographed onSephadex® LH 20, eluting with methanol, and the fractions collected aresubjected to the same treatment as above. 180 mg of the expectedcompound are obtained.

MH⁺ : 674

NMR (DMSO+TFA) 1.00-1.70 : m : 4H; 2.30-3.20 : m : 8H; 4.05 : ds : 4H;4.50 : mt : and 4.80 : mt : 2H; 7.05-8.10 : m : 18H

EXAMPLE 6 ##STR86##

This compound is obtained by working as in Example 5, starting with thecompounds of Preparations 3.9 and 1.2.

MH⁺ : 658 : monochloro isotopic profile

NMR (DMSO+TFA) 1.40-1.70 : m : 4H; 2.40-3.30 : m : 8H; 3.90 : ds : 4H;4.40-4.65 : m : 2H; 6.70-8.00 : m : 15H

EXAMPLE 7 ##STR87##

880 mg of the compound of Preparation 1.2 are dissolved in 15 ml of DMFand 300 μl of Et₃ N, 1.12 g of the compound of Preparation 3.10 areadded and the mixture is left stirring for 5 hours at RT. Afterevaporating the medium to dryness, the residue is taken up in a mixtureof 1N HCl and butanol, and the upper-phase is then separated out aftersettling has taken place and evaporated to dryness. The residue ischromatographed on Sephadex® LH 20, eluting with MeOH. The product thusobtained is taken up in a solution of 1N HCl in butanol and thenevaporated to dryness and the residue is triturated from ether, drained,washed with ether and dried in order to obtain 330 mg of the expectedproduct.

MH⁺ : 730

NMR (DMSO+TFA) ; 1.45-2.25 : m : 4H; 2.30-3.30 : m : 8H; 3.85 : s : and3.95 : s : 4H; 4.50-4.70 : m : 4H; 6.35-8.00 : m : 20H.

EXAMPLE 8 ##STR88##

The debenzylation reaction is carried out according to J. Chem. Educ.,1987, 64, 1062.

425 mg of the compound of Example 7 in 20 ml of methanol are mixed with120 mg of ammonium formate in the presence of 500 mg of 10% Pd/Ccontaining 50% water. After refluxing for 24 hours, the palladium isfiltered off and washed with methanol and the filtrate is evaporated todryness; the residue is dissolved in methanol and then chromatographedon Sephadex® LH 20, eluting with methanol. The product obtained is takenup in a mixture of a solution of 2N HCl in water and butanol; afterstirring, the aqueous fraction is extracted with butanol and thecombined organic fractions are evaporated to dryness. The residue istaken up in ether and the product is filtered off, washed with ether anddried. 171 mg of the expected product are obtained.

MH⁺ : 640

NMR (DMSO+TFA) 1.50-1.85 : m : 4H; 2.40-3.30 : m : 8H; 4.05 : ds : 4H;4.50-4.70 : m : 2H; 6.40-8.20 : m : 15H

The compounds described in Table 8 are prepared as in Examples 7 and 8above.

                                      TABLE 8                                     __________________________________________________________________________     ##STR89##                                                                    Examples                                                                           R.sub.2                                                                                  ##STR90##                                                                              MH.sup.+ NMR (DMSO + TFA)                            __________________________________________________________________________     9                                                                                  ##STR91##                                                                               ##STR92##                                                                              MH.sup.+ : 730  1.45-1.75: m: 4H; 2.40-3.40: m:                               8H; 3.95: s: and 4.00: s: 4H;  4.50-4.70: m: 4H;                              6.40-8.10:  m: 20H                                   10                                                                                  ##STR93##                                                                               ##STR94##                                                                              MH.sup.+ : 640  1.45-180: m: 4H 2.25-3.30: m:                                 8H 3.95: s: 4H 4.45-4.70: m:  2H 6.30: 8.20: m:                               15H                                                  11                                                                                  ##STR95##                                                                               ##STR96##                                                                              MH.sup.+ : 683  1.45-1.80: m: 4H 1.95-2.10: m:                                2H 2.75: ds: 6H 2.50-3.60: m:  12H 4.55: m.t.:                                and 4.70: mt: 2H  6.80-8.10: m:                      __________________________________________________________________________                             16H                                              

EXAMPLE 12 ##STR97##

900 mg of the compound of Preparation 1.2 in 10 ml of DMF and 550 μl ofEt₃ N are mixed with 1.04 g of the compound of Preparation 3.1 and 1.3 gof BOP and a sufficient amount of Et₃ N to reach pH=6-7 are added. Afterstirring for 5 hours, while maintaining the medium at pH=6-7 by additionof Et₃ N, the medium is diluted with ether and is then triturated andthe phases are allowed to separate by settling (twice). The oil ischromatographed on Sephadex® LH 20, eluting with methanol, and theproduct obtained is then chromatographed again on silica, eluting with achloroform/methanol mixture (90/10; v/v). The product obtained is washedwith EtOAc in water, with EtOAc and dried. 313 mg of the expectedproduct are obtained.

MH⁺ : 676 : trichloro isotopic profile

NMR (DMSO+TFA) 1.50-1.85 : m : 4H; 2.70-3.20 : m : 8H; 3.95 : bs : 4H;4.50-4.75 : m : 2H; 6.90-8.00 : m : 11H

Working as in Example 12, the compounds described in the table below areprepared.

                                      TABLE 9                                     __________________________________________________________________________     ##STR98##                                                                    Examples                                                                            R.sub.2 R.sub.3                                                                           ##STR99##                                                                            MH.sup.+  NMR (DMSO + TFA)                           __________________________________________________________________________    13                                                                                   ##STR100##                                                                           H                                                                                 ##STR101##                                                                           MH.sup.+ : 640  1.60-1.80: m: 4H 2.60-3.50: m:                                8H 3.80: s: and 4.00: s: 4H  4.60-4.90: m: 2H                                 6.90-8.70:  m: 15H                                   14                                                                                   ##STR102##                                                                           OH                                                                                ##STR103##                                                                           MH.sup.+ : 640  1.40-1.70: m: 4H 2.50-3.20: m:                                8H 3.85: ds: 4H 4.00: d.d.: 1H  4.30-4.70: m: 2H                              6.55-8.10:  m: 16H                                   __________________________________________________________________________

EXAMPLE 15 ##STR104##

This compound is prepared from the compound prepared in Example 2, stepA, and working as in Example 2, step B, using2-methylpropane-1,2-diamine.

MH⁺ : 652

NMR (DMSO+TFA) 1.30 : s : 6H; 1.30-1.70 : m : 4H; 2.25-3.20 : m : 8H;3.65 : s : 2H; 4.40-4.70 : m : 2H; 6.70-8.05 : m : 16H

EXAMPLE 16 ##STR105##

0.812 g of the compound of Preparation 3.13 and 0.810 g of the compoundof Preparation 1.2 are dissolved in 10 ml of DMF and 1.22 g of BOP areadded, and the mixture is then brought to pH=7 by addition of Et₃ N.After stirring for 18 hours at RT, the medium is evaporated to dryness.The residue is taken up in EtOAc and saturated NaHCO₃ solution, then theaqueous phase is washed with EtOAc and the organic phases are washedwith the KHSO₄ /K₂ SO₄ buffer and with saturated NaCl solution. Theresulting solution is dried over Na₂ SO₄ and evaporated. The residue ischromatographed on silica, eluting with a CHCl₃ /MeOH/concentratedammonia mixture (10/0.5/0.1; v/v/v). 0.390 g of the expected compoundare obtained.

MH⁺ : 625

NMR (DMSO) : 1.5-1.8 : mt : 4H; 2.5-3.7 : m : 8H; 4 : s : 4H; 4.4-4.6 :mt : 1H; 4.7-4.9 : Mt : 1H; 6.8-7 : mt : 3H; 7-7.1 : mt : 2H; 7.4 : d :2H; 7.7-8.1 : m : 5H; 8.3-8.4 : m : 2H; 8.7 : d : 1H; 10.7 : s : 1H.

EXAMPLE 17 ##STR106##

This compound is prepared according to the procedure of the aboveexample, starting with the compound of Preparation 3.14 and that ofPreparation 1.2.

MH⁺ : 625

NMR (DMSO) : 1.4-1.7 : mt : 4H; 2.4-3.3 : m : 4H; 4 : mt : 4H; 4.5-4.7 :mt : 2H; 6.7-6.9 : mt : 2H; 7.2-8.4 : m : 16H; 9 : mt : 1H; 10.7 : s :1H.

EXAMPLE 18 ##STR107##

740 mg of the compound of Preparation 1.2 are placed in 10 ml of DMF and240 μl of DIPEA are added, followed by 740 mg of the compound ofPreparation 3.24. After stirring overnight at RT, ether is added, themixture is decanted and washed again with ether and is then trituratedand decanted. The organic phase is chromatographed on Sephadex® LH 20,eluting with MeOH. The fractions of interest are taken up in a mixturecontaining 5 ml of butanol, 5 ml of 1N HCl and 5 ml of EtOAc. Theorganic phase is washed with 5 ml of 1N HCl and evaporated to dryness.The residue is dissolved in methanol and precipitated with ether. 470 mgof the expected compound are obtained.

MH⁺ : 682

NMR (DMSO) : 1.00 : d : 6H; 1.45-1.80 : m : 4H; 2.30-3.20 : m : 8H; 3.95: ds : 4H; 4.05-4.20 : m : 1H; 4.50-4.70 : m : 2H; 6.30-8.40 : m : 17H(15H aromatic+2H amides); 10.70 : bs : 1H.

Working according to the procedure described above, the compoundsaccording to the invention in the table below are prepared from thecompounds of Preparations 3.25 to 3.32.

                                      TABLE 10                                    __________________________________________________________________________     ##STR108##                                                                   Example                                                                            R.sub.2     NMR              MH.sup.+                                    __________________________________________________________________________    19                                                                                  ##STR109## (DMSO + TFA) 1.50-1.80: m: 4H;  2.25-3.25: m: 8H; 3.40:                       s: 3H; 4.00: ds:  4H; 4.45-4.75: m: 2H; 6.35-6.90: mt:                        4H; 7.30-8.15: m: 11H                                                                          654                                         20                                                                                  ##STR110## (DMSO + TFA) 1.50-1.80: m: 4H;  2.30-3.25: m: 8H; 3.40:                       s: 3H; 4.00: ds:  4H; 4.60: tq: 1H; 6.40: d: 2H; 6.90:                        d:  2H; 7.25-8.05: m: 11H                                                                      654                                         21                                                                                  ##STR111## (DMSO + TFA) 1.50-1.80: m: 4H;  2.35-3.40: m: 8H; 4.00:                       ds: 4H;  4.50-4.70: m: 2H; 6.75-7.00: m: 4H;  7.25-8.10:                      m: 11H           658-660                                     22                                                                                  ##STR112## (DMSO + TFA) 1.40-1.80: m: 4H;  2.40-3.40: m: 8H; 3.95:                       ds: 4H;  4.50-4.65: mt: 1H; 4.75: dq: 1H;  6.90-8.00: m:                      15H              692                                         23                                                                                  ##STR113## (DMSO + TFA) 1.20-1.60: m: 4H;  2.40-3.20: m: 8H; 3.95:                       bs: 4H;  4.25-4.50: mt: 1H; 5.45: t: 1H;  7.00-7.90: m:                       18H              674                                         24                                                                                  ##STR114## (DMSO + TFA) 1.50-1.75: m: 4H;  2.60-3.40: m: 8H; 4.00:                       bs: 4H;  4.50-4.75: m: 2H; 6.55-8.10: m:                                                       642                                         25                                                                                  ##STR115## (DMSO + TFA) 0.60-1.70: m: 15H;  1.80-2.20: m: 2H;                            .2.60-3.40: m:  7H; 3.90: s: 4H; 4.30-4.60: m: 1H;                            7.25-8.25: m: 11H                                                                              630                                         26                                                                                  ##STR116## (DMSO + TFA) 1.50-1.80: m: 4H;  1.90: s: 3H; 2.20-3.30:                       m: 8H; 4.05:  ds: 4H; 4.55-4.75: m: 2H; 6.60-8.10:  m:                        15H              638                                         __________________________________________________________________________

EXAMPLE 27 ##STR117## A)N-(2-(4-Cyanophenyl)-1-(1-pyrrolidinylcarbonyl)ethyl)-3-(4-biphenylyl)-3-(N-Boc)aminopropionamide.

1.19 g of the compound of Preparation 1.1 are placed in 30 ml ofacetonitrile and 0.46 ml of Et₃ N and 1.46 g of the compound ofPreparation 2.17 are added. The mixture is left overnight at RT and isthen evaporated to dryness and the residue is taken up in KHSO₄ /K₂ SO₄and extracted with EtOAc. The extracts are washed with saturated NaClsolution, dried over Na₂ SO₄ and the evaporated to dryness. The expectedcompound crystallizes from heptane and is filtered off, washed withheptane and then dried in order to obtain 1.86 g.

B)N-(2-(4-Cyanophenyl)-1-(1-pyrrolidinylcarbonyl)ethyl)-3-(4-biphenylyl)-3-aminopropionamidetrifluoroacetate.

A mixture containing 1.82 g of the compound of the above step in 15 mlof DCM and 15 ml of TFA is left stirring at RT for 35 minutes. Themixture is evaporated to dryness and the residue is dissolved inisopropanol and evaporated to dryness again. The expected productcrystallizes from Et₂ O and is filtered off, washed with Et₂ O and thendried to give 1.37 g.

C) ##STR118##

1.34 g of the compound of the above step are placed in 25 ml of DCM and850 μl of DIPEA and 522 mg of 2-naphthalenesulphonyl chloride in 5 ml ofDCM are gradually added. After stirring for 2 hours at RT, the mixtureis diluted with 60 ml of DCM and is then washed with KHSO₄ /K₂ SO₄ andwith saturated NaCl solution. The resulting solution is dried over Na₂SO₄ and evaporated to dryness, and the residue is then chromatographedon fine silica, eluting with chloroform containing 1% MeOH. 0.802 g ofthe expected compound is obtained.

NMR (DMSO+TFA) : 1.45-1.85 : m : 4H; 2.40-3.35 : m : 8H; 4.50-4.90 : mt: 2H; 7.05-8.20 : m : 20H.

D)

0.778 g of the compound of the above step is placed in 50 ml ofanhydrous EtOH saturated with HCl, at 0° C.; the mixture is leftstirring in a refrigerator until dissolved and is then left in therefrigerator for 48 hours. It is evaporated to dryness and dried in adesiccator under vacuum in the presence of potassium hydroxide for 2hours. This residue is dissolved in 80 ml of anhydrous EtOH and 206 μlof DIPEA and 93 μl of ethylenediamine are added. After 24 hours at RT,the mixture is evaporated to dryness and the residue is thenchromatographed on Sephadex® LH 20, eluting with methanol. The fractionscontaining the product are taken up in a mixture of 6 ml of butanol and6 ml of 1N HCl. After stirring and separation of the phases by settling,the organic phase is washed with 3 ml of 1N HCl and evaporated undervacuum. The residue is taken up in Et₂ O and the solid formed isfiltered off, washed with Et₂ O and dried to give 0.495 g of theexpected compound.

MH⁺ : 700

NMR : 1.40-1.80 : m : 4H; 2.40-3.30 : m : 8H; 3.80 : s : 2H; 3.95 : s :2H; 4.50-4.80 : mt : 2H; 7.00-8.05 : m : 20H.

EXAMPLE 28 ##STR119##

0.790 g of the compound obtained in Example 2, step A is dissolved in 10ml of EtOH saturated with HCl, at 0° C., and the mixture is leftstirring for 48 hours at 0° C. It is evaporated to dryness and theresidue is then taken up in EtOH and evaporated (twice). The residue istaken up in DCM and evaporated (twice). The resulting residue is takenup in 20 ml of EtOH and then neutralized by addition of Et₃ N. 0.109 mlof diaminopropane is added and the mixture is left for 24 hours at RT.It is evaporated to dryness and the residue is taken up in DCM andwashed with KHSO₄ /K₂ SO₄. Chromatography on silica is then carried out,eluting with a chloroform/methanol/concentrated ammonia mixture (10/1/1;v/v/v). 0.12 g of the expected compound is obtained.

MH⁺ : 638

EXAMPLE 29 ##STR120##

0.888 g of the compound of Preparation 3.2 and 0.957 g of the compoundof Preparation 1.4 are dissolved in 5 ml of DMF; 1.22 g of BOP are addedand the medium is then adjusted to pH=7 by addition of Et₃ N. Afterstirring for 18 hours at RT, the mixture is evaporated to dryness andthe residue is then taken up. in EtOAc. The solution is washed withNaHCO₃ solution, with KHSO₄ /K₂ SO₄ and with saturated NaCl solution andis then dried over Na₂ SO₄ and evaporated. The residue is taken up inether and a few drops of hexane and is then filtered and dried over Na₂SO₄ in order to obtain 1.1 g of the expected compound.

B)

1 g of the compound of the above step is dissolved in 20 ml of EtOHsaturated with HCl, at 0° C. After 48 hours at +4° C., the solution isevaporated and the residue is taken up in EtOH (3 times) and is thenevaporated and taken up in DCM (twice). The residue is taken up in 20 mlof EtOH, the pH is then adjusted to 7 by addition of Et₃ N and 0.122 mlof ethylenediamine is added. After stirring for 18 hours at RT, themixture is evaporated to dryness and is then taken up in DCM and washedwith 10 ml of KHSO₄ /K₂ SO₄. This solution is chromatographed on silica,eluting with a chloroform/methanol/aqueous ammonia mixture (10/1/0.1;v/v/v) in order to obtain 0.640 g of the expected compound.

MH⁺ : 650

NMR (DMSO+TFA) : 0.8-1.2 : m : 4H; 2.3-3.1 : m : 4H; 4 : s : 4H; 4.3-4.7: m : 4H; 5.4-5.7 : mt : 2H; 6.8-7.9 : m : 15H; 8.2 : d : 1H.

Working as in Example 12, the compounds described in Table 11 below areprepared.

                  TABLE 11                                                        ______________________________________                                         ##STR121##                                                                   Example R.sub.1        R.sub.2     MH.sup.+                                   ______________________________________                                        30                                                                                     ##STR122##                                                                                   ##STR123## 642-644                                    31                                                                                     ##STR124##                                                                                   ##STR125## 628                                        ______________________________________                                    

Working according to the procedure of Example 2, steps A and B, thecompounds according to the invention in the table below are prepared.

                  TABLE 12                                                        ______________________________________                                         ##STR126##                                                                   Example                                                                              --NR.sub.4 R.sub.5                                                                         m.p. ° C./NMR (DMSO + TFA)                                                               MH.sup.+                                ______________________________________                                        32                                                                                    ##STR127##  170° C.  2.40-3.45:m:12H; 4.00:se:4H;                                  4.60-4.90:m:2H;  6.80-8.15:m:16H                                                                640                                     33                                                                                    ##STR128##  0.6-0.9:mt:6H; 2.2-3:m:7H;  3.9:s:4H; 4.3-4.9:m:3H;                           6.7-7.8:m:15H; 8.:d:1H                                                                          626                                     34                                                                                    ##STR129##  0.6-1.6:m:8H; 1.6-1.9:mt:2H;  2.4-3.1:m:4H;                                   3.2-4:m:6H;  4.2-4.8:m:2H; 6.8-7.9:m:15H;                                                       652:1H                                  ______________________________________                                    

EXAMPLE 35 ##STR130##

580 mg of the compound obtained in Example 2, step A are mixed with 250mg of paraformaldehyde and 60 mg of para-toluenesulphonic acidmonohydrate in 25 ml of benzene. The mixture is refluxed in Dean-Starkapparatus for 1 hour. The reaction medium is then washed with saturatedNaHCO₃ solution and with saturated NaCl solution and then dried over Na₂SO₄ and evaporated to dryness. The residue is chromatographed on finesilica, eluting with chloroform, in order to obtain 165 mg of theexpected compound.

NMR (DMSO+TFA) : 1.55-1.85 : m : 4H; 2.40-3.40 : m : 8H; 4.60-5.50 : m :4H; 6.90-8.30 : m : 16H.

B)

The process is performed as in Example 2, step B in order to obtain theexpected compound, which is purified by chromatography on Sephadex® LH20, eluting with MeOH.

MH⁺ : 636

NMR (DMSO+TFA) : 1.50-1.80 : m : 4H; 2.40-3.30 m : 8H; 3.95 : s : 4H;4.90-5.45 : m : 4H; 6.90-8.10 : m : 16H.

EXAMPLE 36 ##STR131##

105 g of the compound of Example 2, step A are mixed with 0.262 g ofpotassium carbonate in 12 ml of DMF and 423 μl of methyl iodide areadded. The following day, the mixture is evaporated to dryness and theresidue is taken up in water and EtOAc. The organic phase is washed withwater and with saturated NaCl solution and is then dried over Na₂SO₄.0.6 g of the expected compound is obtained.

MH⁺ : 595

NMR (DMSO+TFA) : 1.50-1.80 : m : 4H; 2.20-3.30 : m : 8H; 2.55 : bs : 3H;4.35-4.65 : mt : 1H; 5.45-5.60 : mt : 1H; 7.10-8.45 : m : 16H.

B)

The process is performed as in Example 2, step B, in order to obtain theexpected compound, which is purified by chromatography on silica,eluting with DCM/MeOH (93/7; v/v) and then by a second chromatography onSephadex® LH 20, eluting with methanol, m.p.=154° C.

MH⁺ : 638

NMR (DMSO+TFA) : 1.20-1.80 : m : 4H; 2.10-2.40 : mt : 2H; 2.50 : bs :3H; 2.40-3.20 : m : 6H; 4.00 : s : 4H; 4.40-4.70 : mt : 1H; 5.35-5.55 :mt : 1H; 7.05-8.40 : m : 16H.

EXAMPLE 37 ##STR132##

A mixture containing 0.79 g of the compound of Preparation 3.35, 0.79 gof the compound of Preparation 1.1 and 1.06 g of BOP in 10 ml of DMF isprepared, DIPEA is added in order to obtain pH =7 and the mixture isleft stirring for 2 hours at RT. The mixture is extracted with EtOAc andthe extracts are washed with H₂ O, with 0.25N NaOH solution, with 0.25NHCl solution, with water and then with saturated NaCl solution. Theresulting solution is dried over Na₂ SO₄ and then evaporated and theresidue is chromatographed on silica, eluting with a chloroform/MeOHmixture (95/5; v/v).

B)

The expected product is obtained by working as in Example 2, step B. Itis purified by chromatography on Sephadex® LH 20, eluting with DCM/MeOH(3/2; v/v).

MH⁺ : 636

NMR (DMSO+TFA) : 1.40-1.80 : m : 4H; 2.60-3.60 : mt : 9H; 3.95 : s : 4H;4.60-5.20 : m : 2H; 6.20-8.50 : m : 15H.

EXAMPLE 38 ##STR133##

533 g of the compound of Preparation 3.37 are placed in 10 ml of DMF and205 μl of DIPEA, 573 mg of the compound of Preparation 1.2 are added andthe mixture is then left overnight at RT. This mixture is diluted with100 ml of Et₂ O and the gum-formed is then chromatographed on Sephadex®LH 20, eluting with MeOH. The fractions of interest are combined andfiltered and the residue is taken up in 6 ml of butanol and 6 ml of HCl.The organic phase is separated out after settling of the phases hastaken place and is then washed with 3 ml of 1N HCl and evaporated todryness. The product crystallizes from Et₂ O and is filtered off anddried to give 430 mg of the expected compound.

MH⁺ : 650

NMR (DMSO+TFA) : 1.50-1.90 : m : 4H; 2.50-3.80 : m : 12H; 4.00 : ds :4H; 4.45-4.80 : m : 1H; 5.40 : dq : 1H; 6.75-8.35 : m : 15H.

EXAMPLE 39 ##STR134##

A reaction medium containing 710 mg of the compound of Preparation 3.2,750 mg of the compound of Preparation 1.8 and 0.98 g of BOP in 12 ml ofDMF is stirred and DIPEA is added to bring the mixture to pH=6. Afterstirring for 2 hours at RT, the mixture is extracted with EtOAc. Thecrude product obtained is chromatographed on silica, eluting withEtOAc/toluene (3/2; v/v). 0.52 g of the expected compound is obtained inthe form of a white solid.

B)

The expected compound is obtained by working as in Example 2, step B. Itis purified on Sephadex® LH 20, eluting with DCM/MeOH (3/2; v/v).

MH⁺ : 638

NMR (DMSO+TFA) : 1.40-1.75 : m : 4H; 2.30-3.20 : m : 11H; 4.00 : s : 4H;4.60-5.40 : mt : 2H; 6.80-7.15 : m : 16H.

EXAMPLE 40 ##STR135## A)

0.42 g of the compound of Example 2, step A is treated with 10 ml ofsaturated hydrochloric ethanol at -10° C. After 72 hours at +4° C., themixture is evaporated to dryness and is then dried under vacuum in thepresence of potassium hydroxide. 0.51 g of unpurified product isobtained in the form of the hydrochloride.

B)

0.5 g of the ethyl imidate obtained in the above step is cooled to 0° C.and 10 ml of saturated ammoniacal ethanol are added, at 0° C. Themixture is allowed to return to RT. The following day, the mixture isevaporated to dryness and the residue is then chromatographed on silica,eluting with a DCM/MeO mixture (90/10; v/v), then another chromatographyon Sephadex® LH 20 is carried out, eluting with MeOH. 0.15 g of theexpected compound is obtained, m.p.=180° C.

MH⁺ : 598

NMR (DMSO) : 1.40-1.70 : m : 4H; 2.25-3.10 : m : 8H; 4.40-4.70 : m : 2H;6.80-8.30 : m : 18H.

EXAMPLE 41 ##STR136## A) 4-Aminobutyronitrile.

This compound is prepared according to J. Am. Soc., 1952, 74, 1836. 6.7ml of 4-bromobutyronitrile and 35 ml of liquid ammonia are placed in abomb at -50° C. After closing, the bomb is left at RT for 48 hours. Theresidue is taken up in 50% NaOH solution and is then extracted withether; the organic phase is dried over Na₂ SO₄, evaporated to drynessand then chromatographed on silica, eluting with DCM/MeOH/NH₄ OH(90/10/0.3; v/v/v). 1.07 g of the expected compound are obtained.

B)

0.62 g of the compound of Example 2, step A is treated with 10 ml ofsaturated hydrochloric ethanol at -10° C. After 24 hours at +4° C., themixture is evaporated to dryness and the residue is then dried undervacuum. 0.8 g of unpurified product is obtained in the form of thehydrochloride.

C) ##STR137##

0.8 g of the compound prepared in the above step is dissolved in 10 mlof EtOH and 108 mg of 4-aminobutyronitrile diluted in 10 ml of EtOH areadded dropwise. The following day, the mixture is evaporated to drynessand the residue is taken up in MeOH and a few drops of hydrochloricether are then added. After evaporating to dryness, the residue ischromatographed on silica, eluting with DCM/MeOH (90/10; v/v) in orderto obtain 0.38 g of the expected compound, m.p.=144° C.

MH⁺ : 665

D)

10 ml of saturated hydrochloric ethanol at -10° C. are added to 0.35 gof the compound obtained in the above step, at 0° C. The mixture is leftin a refrigerator overnight. The following day, the mixture isevaporated to dryness and the residue is dried under vacuum in thepresence of KOH. The product obtained (0.35 g) is dissolved in 80 ml ofethanol; 43 μl of ethylenediamine in 10 ml of ethanol are addeddropwise, After stirring overnight, the mixture is evaporated to drynessand the residue is then chromatographed on silica, eluting with aDCM/MeOH mixture (50/50; v/v). 0.075 g of the expected compound isobtained, m.p.=195° C.

MH⁺ : 708

NMR (DMSO+TFA) : 1.35-1.50 : m : 4H; 1.80-2.00 : mt : 2H; 2.40-3.20 : m: 10H; 3.40 : mt : 2H; 3.70 : s : 4H; 4.40-4.70 : mt : 2H; 6.70-8.05 : m: 16H.

EXAMPLE 42 ##STR138##

This compound is prepared according to the process described in Example12, starting with the compounds of Preparations 1.9 and 3.2.

MH⁺ : 626

EXAMPLE 43 ##STR139## (R,R) isomer. A) ##STR140## (R,R) isomer.

1.44 g of the compound of Preparation 1.13 in 40 ml of acetonitrile aremixed with 700 μl of DIPEA and 1.81 g of the compound of Preparation3.39. The precipitate formed is dissolved in DCM and is then washed withKHSO₄ /K₂ SO₄, with saturated NaHCO₃ solution and with saturated NaClsolution. The resulting solution is dried and evaporated to dryness, andthe product obtained (1.57 g) is used without further purification inthe following step.

NMR (DMSO) : 1.40-1.60 : m : 4H; 2.20-3.10 : m : 8H; 4.40 : dq : 1H;4.60 : dq : 1H; 6.80-8.35 : m : 18H.

B)

1 g of the product of the above step is suspended in 30 ml ofHCl-saturated anhydrous ethanol at 0° C. and the mixture is leftstirring for 24 hours in a refrigerator. The mixture is evaporated todryness and dried in a desiccator in the presence of potassiumhydroxide. The product is placed in 100 ml of anhydrous ethanol and 273μl of DIPEA and 136 μl of ethylenediamine are added. After stirring for24 hours at RT, the mixture is evaporated to dryness and the residue istaken up in a butanol/chloroform/1N HCl mixture (1/1/1; v/v/v). Thephases are separated out after settling has taken place and the organicphase is then washed with 1N HCl and evaporated to dryness. The residueis chromatographed on Sephadex® LH 20, eluting with MeOH. The productobtained is treated with a butanol/chloroform/1N HCl mixture (1/1/1;v/v/v) and then concentrated. After crystallization from Et₂ O, thecrystals are filtered off, washed with Et₂ O and then dried. 198 mg ofthe expected compound are obtained.

α_(D) ²⁵ =+38.1° (c=1, DMF)

MH⁺ : 624

NMR (DMSO) ; 1.50-1.75 : m : 4H; 2.40-2.70 : mt : 2H; 2.75-3.25 : m :6H; 4.05 : s : 4H; 4.55 : dq : 1H; 4.75 : dq : 1H; 6.95-8.20 : m : 16H;8.50 : t : 2H; 10.80 : bs : 2H.

Using the procedure described in Example 43 and in the correspondingpreparations, the various isomers described in the table below areprepared.

                  TABLE 13                                                        ______________________________________                                         ##STR141##                                                                                       α.sub.D.sup.25 = (c = 1, DMF)                       Example  Isomer     m.p. ° C.                                                                            MH.sup.+                                    ______________________________________                                        44       (S,S)      -41.9° 624                                         45       (R,S)      +42.2° 624                                         46       (S,R)      m.p. = 195-201° C.                                                                   624                                         ______________________________________                                    

NMR (DMSO) of Example 44 : 1.50-1.75 : m : 4H; 2.40-2.70 : mt : 2H;2.75-3.25 : m : 6H; 4.05 : s : 4H; 4.55 : dq : 1H; 4.75 : dq : 1H;6.95-8.20 : m : 16H; 8.50 : t : 2H; 10.80 : bs : 2H.

NMR (DMSO) of Example 45 : 1.65-1.90 : m: 4H; 2.40-3.45 : m : 8H; 4.05 :s : 4H; 4.55-4.85 : mt : 2H; 6.95-8.25 : m : 16H; 8.45 : t : 2H; 10.80 :bs : 2H.

NMR (DMSO+TFA) of Example 46 : 1.50-1.80 : m : 4H; 2.25-2.95 : m : 4H;3.00-3.40 : m : 4H; 4.00 : s : 4H; 4.30-4.75 : mt : 2H; 6.80-8.15 : m :16H.

EXAMPLE 47 ##STR142## (R,R) isomer. ##STR143## (R,R) isomer.

These 2 compounds are prepared in 2 successive steps according to theprocedure described in Example 27, steps C and D, from the compoundobtained in Preparation 4.2 and (2,4,6-trichlorophenyl)sulphonylchloride.

I : α_(D) ²⁵ =-24° (c=0.5; MeOH)

MH⁺ : 676 with trichloroisotopic profile

NMR (DMSO) : 1.50-1.65 : m : 4H; 2.45-3.20 : m : 8H; 3.95 : s : 4H;4.40-4.70 : mt : 2H; 7.05 : s : 5H; 7.45 : d : 2H; 7.50 : s : 2H; 7.90 :d : 2H; 8.45 : d : 1H; 8.80 : d : 1H; 10.70 : bs : 1H.

II : NMR (DMSO): 1.50-1.60 : m : 4H; 2.45-3.15 : m : 8H; 4.40-4.55 : dq: 1H; 4.60-4.75 : dq : 1H; 7.05 : s : 5H; 7.40 : d : 2H; 7.50 : s : 2H;7.70 : d : 2H; 8.40 : d : 1H; 8.70 : d : 1H.

EXAMPLE 48 ##STR144## (R,R) isomer ##STR145## (R,R) isomer

This compound is prepared in 2 steps according to the proceduredescribed in Example 43, starting with the compounds of Preparations3.43 and 1.13.

I : α_(D).sup.≅ =+57° (c=1; DMF)

MH⁺ : 658 and 660

NMR (DMSO+TFA) : 1.40-1.60 : m : 4H; 2.25-3.10 : m : 8H; 3.95 : s : 4H;4.45-4.65 : mt : 2H; 6.80-8.05 : m : 15H.

II : α_(D) ²⁵ =+61° (c=1; DMF)

NMR (DMSO) : 1.40-1.60 : m : 4H; 2.25-3.10 : m : 8H; 4.35-4.60 : mt :2H; 6.85-8.05 : m : 15H; 8.25 : t : 2H.

EXAMPLE 49 ##STR146## (S), (R,S) isomer.

800 mg of the compound of Preparation 1.2 in 20 ml of DMF are mixed with250 μl of DIPEA and 933 mg of the compound of Preparation 3.45. Afterstirring overnight at RT, the mixture is diluted with ether and theproduct is then isolated by decanting. The gum formed is triturated fromether and is then chromatographed on Sephadex® LH 20, eluting with MeOH.The fractions of interest are evaporated and the residue is taken up ina butanol/EtOAc/1N HCl mixture (1/1/1; v/v/v). The organic phase isseparated out after settling has taken place and evaporated to dryness,the residue is taken up in Et₂ O and the product is filtered off anddried to give 770 mg of the expected compound.

MH⁺ : 638

NMR (DMSO+TFA) : 1.55-1.85 : m : 4H; 2.05-3.50 : m : 11H; 3.75 : s : 2H;4.00 : s : 2H; 4.45-4.95 : mt : 1H; 6.70-8.20 : m : 16H.

EXAMPLE 50 ##STR147##

This compound is prepared from the compound of Preparation 3.46 and fromthe compound of Preparation 1.2 according to the procedure of Example 5.

NMR (DMSO+TFA) : 1.5-1.8 : mt : 6H; 2.7-3.6 : m : 6H; 3.6 : mt : 3H;3.6-3.9 : mt : 8H; 4.6-4.8 : mt : 2H; 6.5 : d : 2H; 6.8-7 : m : 4H;7.4-8.1 : m : 8H; 8.4 : d : 2H.

EXAMPLE 51 ##STR148##

0.4 g of the compound obtained in the above example is dissolved in 8 mlof TFA and 0.2 ml of thioanisole. After stirring for 24 hours at RT, themixture is evaporated, the residue is then taken up in Et₂ O and theproduct is filtered off. It is washed several times with ether and thendried over Na₂ SO₄ in order to obtain 0.385 g of the expected compound.

MH⁺ : 700

NMR (DMSO+TFA) : 1.7-2.1 : mt : 6H; 2.5 : d : 2H; 2.9-3.7 : m : 13H;3.7-4 : m : 13H; 4.8 : mt : 1H; 5.3 : mt : 1H; 7.2-8.3 : m : 10H; 8.5 :s : 1H.

EXAMPLE 52 ##STR149##

The compound obtained in the above example is dissolved in 50 ml of MeOHcontaining 0.106 g of KOH. After stirring for 18 hours at RT, the mediumis acidified to pH=2 by addition of a saturated solution of HCl gas indioxane. This mixture is evaporated and the residue is taken up in 5 mlof water and triturated. The product is filtered off, washed with waterand then dried in order to obtain 0.160 g of the expected compound.

MH⁺ =604

EXAMPLE 53 ##STR150## (R,R) isomer

This compound is prepared according to the procedure described inExample 43, but using the 1-naphthalenesulphonyl chloride in Preparation3.38.

α_(D) ²⁵ =-18° (c=1; DMF)

MH⁺ : 624

NMR (DMSO) : 1.50-1.75 : m : 4H; 2.25-3.20 : m : 8H; 4.05 : s : 4H;4.50-4.75 : mt : 2H; 6.85-8.70 : m : 18H (16H aromatic+2H).

EXAMPLE 54 ##STR151##

This product is prepared in 2 steps according to the procedure describedfor Example 2, using the compounds obtained from Preparations 3.2 and1.14.

MH⁺ : 638

NMR (DMSO+TFA) : 0.95 : ds : 3H; 1.30-1.60 : m : 4H; 2.40-3.40 : m : 8H;4.00 : ds : 4H; 4.75 : mt : 1H; 6.65-8.15 : m : 16H.

EXAMPLE 55 ##STR152## (R,R) isomer

1.30 g of the product obtained according to the procedure of Example 43,step A are stirred for 18 hours at 4° C. in 45 ml of methanol containing22 g of HCl gas. The reaction medium is concentrated under vacuum,re-evaporated twice with MeOH and the residue is dried under vacuum inthe presence of KOH. This product is dissolved in 250 ml of methanol and0.18 ml of ethylenediamine diluted in 15 ml of methanol are added over90 minutes, after which the mixture is stirred for a further 18 hours.The reaction medium is concentrated under vacuum to 10 ml, an Et₂ O/HClsolution is added in order to bring the mixture to pH=3 and theresulting mixture is concentrated to dryness. The residue is purified bypartition chromatography on Sephadex® G 25, using the nBuOH/iPrOH/H₂ Osolvent system (4/0.2/5; v/v/v). The product with the cyclized amidine(410 m g) as described in Example 43 is first isolated, followed by theexpected product with the open amidine (410 mg).

α_(D) ²⁵ =+37.6° (c=0.5; DMF)

NMR (DMSO+TFA) : 1.50-1.70 : m : 4H; 2.30-3.15 : m : 10H; 3.70 : t : 2H;4.55 : t : 1H; 4.70 t : 1H; 6.90-8.10 : m : 16H.

EXAMPLE 56

Gelatin capsule containing a 10 mg dose

    ______________________________________                                        Compound of Example 43 (weight expressed as                                                           10.0 mg                                               equivalent in non-salified form)                                              Lactose monohydrate 200 mesh                                                                          qs                                                    Methylhydroxypropylcellulose 6 mPa · s                                                       3.0 mg                                                Crosslinked sodium carboxymethylcellulose                                                             4.5 mg                                                Magnesium stearate      1.5 mg                                                Purified water for wet granulation                                            ______________________________________                                    

For a No. 3 size "opaque-white" gelatin capsule filled to 150 mg.

EXAMPLE 57

Splittable uncoated tablet containing a 50 mg dose

    ______________________________________                                        Compound of Example 43 (weight expressed as                                                           50.0 mg                                               equivalent in non-salified form)                                              Lactose monohydrate 200 mesh                                                                          qs                                                    Microcrystalline cellulose 50 μm                                                                   27.0 mg                                               Methylhydroxypropylcellulose 6 mPa · s                                                       3.6 mg                                                Crosslinked sodium carboxymethylcellulose                                                             5.4 mg                                                Magnesium stearate      1.8 mg                                                Purified water for wet granulation                                            ______________________________________                                    

For a finished splittable uncoated tablet containing ;80 mg

EXAMPLE 58

Gelatin capsule containing a 1 mg dose

    ______________________________________                                        Compound of Example 43 (weight expressed as                                                           1.0 mg                                                equivalent in non-salified form)                                              Lactose monohydrate 200 mesh                                                                          qs                                                    Methylhydroxypropylcellulose 6 mPa · s                                                       3.0 mg                                                Crosslinked sodium carboxymethylcellulose                                                             4.5 mg                                                Magnesium stearate      1.5 mg                                                Purified water for wet granulation                                            ______________________________________                                    

For a NO. 3 size "opaque-white" gelatin capsule filled to 100 mg.

What is claimed is:
 1. A compound of formula: ##STR153## in which: R₁ isa quinolyl or an isoquinolyl, the said rings being unsubstituted orsubstituted one or more times with R₁₀ ;R₂ is a phenyl which isunsubstituted or substituted one or more times with R₁₁, a phenyl(C₁-C₄)alkyl which is unsubstituted or substituted one or more times on thephenyl with R₁₁, a naphthyl which is unsubstituted or substituted one ormore times with R₁₁, a cyclohexyl which is unsubstituted or substitutedone or more times with R₁₁ ; or R₂ and R₉ are linked together andconstitute a (C₃ -C₅)alkylene which is unsubstituted or substituted withR₁₂ or a (C₂ -C₄)alkylene which is interrupted with an oxygen atom or asulphur atom and is unsubstituted or substituted with R₁₂ ; or R₂ andR₉, together with the carbon atom and the nitrogen atom to which theyare attached, constitute tetrahydroisoquinoline which is unsubstitutedor substituted one or more times with a halogen, a hydroxyl, a (C₁-C₄)alkyl, a (C₁ -C₄)alkoxy or a benzyloxy; R₃ is hydrogen or ahydroxyl; R₄ and R₅ are each independently hydrogen or a (C₁ -C₄)alkyl;or R₄ and R₅, together with the nitrogen atom to which they areattached, constitute a heterocyclic radical chosen from 1-pyrrolidinyl,1-piperidyl, perhydro-1-azepinyl, 4-morpholinyl, 4-oxo-1-piperidyl,dihydro-1-pyrrolyl or dihydro-2-imidazolyl, the said heterocyclicradicals being unsubstituted or substituted one or more times with R₁₃ ;R₆ is hydrogen and R₆ can also be R₈ when R₇ is hydrogen; R₇ is hydrogenor a (C₁ -C₄)alkyl; R₈ is hydrogen; a benzyl which is unsubstituted orsubstituted on the phenyl one or more times with R₁₃ ; or a group ZR₁₄ ;or R₇ and R₈, together with the nitrogen atom to which they areattached, constitute a heterocyclic radical chosen from 1-pyrrolidinyl,1-piperidyl, 1-perhydro-1-azepinyl, 4-morpholinyl,tetrahydro-2-pyrimidinyl, 1-piperazinyl or 1-piperazinyl substituted inposition 4 with a (C₁ -C₄)alkyl or a benzyl; or, when R₇ is hydrogen, R₆and R₈ are linked together to form a (C₂ -C₄)alkylene which isunsubstituted or substituted one or more times with a (C₁ -C₄)alkyl; R₉is hydrogen, a (C₁ -C₄)alkyl or a phenyl(C₁ -C₄)alkyl which isunsubstituted or substituted on the phenyl one or more times with R₁₁ ;R₁₀ is a halogen, a (C₁ -C₄)alkyl, a (C₁ -C₄)alkoxy, a hydroxyl, anamino, a (C₁ -C₄)alkylamino or a di(C₁ -C₄)alkylamino; R₁₁ is a halogen,a (C₁ -C₄)alkyl, a trifluoromethyl, a phenyl, a hydroxyl, a (C₁-C₄)alkoxy or a benzyloxy; or R₁₁ is in the ortho position to the phenylrepresenting R₂ and forms with R₃ a methylene group or an ethylenegroup; or R₁₁ is in the ortho position to the phenyl representing R₂ andforms with R₉ a methylene group or an ethylene group; R₁₂ is a halogen,a (C₁ -C₄)alkyl, a hydroxyl, a (C₁ -C₄)alkoxy, a benzyloxy, an oxo, aphenyl, an acetyloxy or a trifluoroacetyloxy; R₁₃ is a (C₁ -C₄)alkyl, ahalogen or a hydroxyl; R₁₄ is a methyl, an amino, a (C₁ -C₄)alkylamino,a di(C₁ -C₄)alkylamino, a tri(C₁ -C₄)alkylammonium, an amidino, a (C₁-C₄)alkylamidino, a guanidino, a (C₁ -C₄)alkylguanidino, a hydroxyl, a(C₁ -C₄)alkoxy, a (C₁ -C₄)alkoxycarbonyl, a group--AlkN(R₁₅)Alk'N(R₁₅)₂, or a heterocyclic radical chosen from1-pyrrolidinyl, 1-piperidyl, perhydro-1-azepinyl, pyridyl, imidazolyl,dihydroimidazolyl, inidazolidinyl, pyrimidinyl and indolyl; R₁₅ and R'₁₅are, independently of each other, hydrogen or a (C₁ -C₄)alkyl; R₁₆ ishydrogen or a methyl, or R₁₆ forms with R₉ a methylene group; R₁₇ ishydrogen or a methyl; Alk and Alk' are, independently of each other, a(C₁ -C₄)alkylene; Z is a (C₂ -C₁₂)alkylene or a (C₁ -C₆)alkylene whichis interrupted or substituted with a (C₅ -C₇)cycloalkyl or with aphenyl; C* is an asymmetric carbon atom;as well as the salts thereofwith inorganic or organic acids.
 2. A compound of formula (I) accordingto claim 1, in which:R₁ is a quinolyl or an isoquinolyl, the said ringsbeing unsubstituted or substituted one or more times with R₁₀ ; R₂ is aphenyl which is unsubstituted or substituted one or more times with R₁₁,a phenyl(C₁ -C₄)alkyl which is unsubstituted or substituted one or moretimes on the phenyl with R₁₁, or a naphthyl which is unsubstituted orsubstituted one or more times with R₁₁ ; or R₂ and R₉ are linkedtogether and constitute a (C₃ -C₅)alkylene which is unsubstituted orsubstituted with R₁₂ or a (C₂ -C₄)alkylene which is interrupted with anoxygen atom or a sulphur atom and is unsubstituted or substituted withR₁₂ ; or R₂ and R₉ together with the carbon atom and the nitrogen atomto which they are attached, constitute tetrahydroisoquinoline which isunsubstituted or substituted one or more times with a halogen, ahydroxyl, a (C₁ -C₄)alkyl, a (C₁ -C₄)alkoxy or a benzyloxy; R₃ ishydrogen or a hydroxyl; R₄ and R₅ are each independently hydrogen or a(C₁ -C₄)alkyl; or R₄ and R₅, together with the nitrogen atom to whichthey are attached, constitute a heterocyclic radical chosen from1-pyrrolidinyl, 1-piperidyl, perhydro-1-azepinyl, 4-morpholinyl or4-oxo-1-piperidyl, the said heterocyclic radicals being unsubstituted orsubstituted with R₁₃ ; R₆ is hydrogen, R₆ can also be R₈ when R₇ ishydrogen; R₇ is hydrogen or a (C₁ -C₄)alkyl; R₈ is hydrogen; a benzylwhich is unsubstituted or substituted on the phenyl one or more timeswith R₁₃ ; or a group ZR₁₄ ; or R₇ and R₈, together with the nitrogenatom to which they are attached, constitute a heterocyclic radicalchosen from 1-pyrrolidinyl, 1-piperidyl, 1-perhydro-1-azepinyl,4-morpholinyl, 1-piperazinyl or 1-piperazinyl substituted in position 4with a (C₁ -C₄)alkyl or a benzyl; or, when R₇ is hydrogen, R₆ and R₈ arelinked together to form a (C₂ -C₄)alkylene which is unsubstituted orsubstituted one or more times with a (C₁ -C₄)alkyl; R₉ is hydrogen, a(C₁ -C₄)alkyl or a phenyl(C₁ -C₄)alkyl which is unsubstituted orsubstituted on the phenyl one or more times with R₁₁ ; R₁₀ is a halogen,a (C₁ -C₄)alkyl, a (C₁ -C₄)alkoxy, a hydroxyl, an amino, a (C₁-C₄)alkylamino or a di(C₁ -C₄)alkylamino; R₁₁ is a halogen, a (C₁-C₄)alkyl, a hydroxyl, a (C₁ -C₄)alkoxy or a benzyloxy; R₁₂ is ahalogen, a (C₁ -C₄)alkyl, a hydroxyl, a (C₁ -C₄)alkoxy, a benzyloxy, anoxo or a phenyl; R₁₃ is a (C₁ -C₄)alkyl, a halogen or a hydroxyl; R₁₄ isa methyl, an amino, a (C₁ -C₄)alkylamino, a di(C₁ -C₄)alkylamino, atri(C₁ -C₄)alkylammonium, an amidino, a (C₁ -C₄)alkylamidino, aguanidino, a (C₁ -C₄)alkylguanidino, a hydroxyl, a (C₁ -C₄)alkoxy, a (C₁-C₄)alkoxycarbonyl, a group --AlkN(R₁₅)Alk'N(R'₁₅)₂, or a heterocyclicradical chosen from 1-pyrrolidinyl, 1-piperidyl, perhydro-1-azepinyl,pyridyl, imidazolyl, dihydroimidazolyl, imidazolidinyl, pyrimidinyl andindolyl; R₁₅ and R'₁₅ are, independently of each other, hydrogen or a(C₁ -C₄)alkyl; R₁₆ is hydrogen; R₁₇ is hydrogen; Alk and Alk' are,independently of each other, a (C₁ -C₄)alkylene; Z is a (C₂-C₁₂)alkylene or a (C₁ -C₆)alkylene which is interrupted or substitutedwith a (C₅ -C₇)cycloalkyl or with a phenyl;as well as the salts thereofwith inorganic or organic acids.
 3. A compound of formula (I) accordingto claim 1 which satisfies at least one of the followingconditions:a--R₁ is a quinolyl; R₂, R₃, R₄, R₅, R₆, R₇, R₈, R₉, R₁₆ andR₁₇ being as defined in claim 1; b--R₂ is a phenyl which isunsubstituted or substituted with R₁₁ ; R₁, R₃, R₄, R₅, R₆, R₇, R₈, R₉,R₁₆ and R₁₇ being as defined in claim 1; c--NR₄ R₅ is a 1-pyrrolidinylgroup; R₁, R₂, R₃, R₆, R₇, R₈, R₉, R₁₆ and R₁₇ being as defined in claim1; d--C(NR₆)NR₇ R₈ is a 4,5-dihydro-2-imidazolyl; R₁, R₂, R₃, R₄, R₅,R₉, R₁₆ and R₁₇ being as defined in claim 1; e--R₃ =R₉ =R₁₆ =R₁₇ =H; R₁,R₂, R₄, R₅, R₆, R₇ and R₈ being as defined in claim 1;and the saltsthereof with inorganic or organic acids.
 4. A compound according toclaim 3 of formula: ##STR154## in which: R_(2a) is a phenyl which isunsubstituted or substituted in a meta or para position with R₁₁ ; a1-naphthyl or a 2-naphthyl;R₁, R₆, R₇, R₈ and R₁₁ are as defined inclaim 1 for (I);and the salts thereof with inorganic or organic acids.5. A compound according to claim 4 of formula: ##STR155## in which:R_(1a) is a 2-quinolyl;R_(2a) is as defined in claim 4 for (Ia);and thesalts thereof with inorganic or organic acids.
 6. A compound of formula(I'a) according to claim 5, in which R_(2a) is a phenyl which isunsubstituted or substituted in a meta position or para position withR₁₁.
 7. A compound according to claim 1 having (R,R) isomerism on theC*-labelled carbon atoms.
 8. A process for the preparation of thecompounds of formula (I) according to claim 1 wherein:a1) a compound offormula: ##STR156## in which R₁, R₂, R₃, R₄, R₅, R₉, R₁₆, R₁₇ and C*have the definitions given in claim 1 for (I), in the form of a pureenantiomer or a mixture of isomers in any proportion, is treated with analcohol of formula R--OH in which R is a (C₁ -C₄)alkyl, in acidicmedium, in order to form an intermediate imidate which is reacted withan amine of formula HNR₇ R₈ (III) or a diamine of formula H₂ NR₆ R₈ NH₂(IV) in which R₆, R₇ and R₈ have the definitions given in claim 1 for(I); b1) the compound of formula (I) thus obtained is isolated in baseform or salt form, c1) where appropriate, another salt of the compoundof formula (I) is prepared.
 9. A process for the preparation of acompound of formula (I) according to claim 1 wherein:a2) a compound offormula: ##STR157## In which X is hydrogen or a Boc group and R₄, R₅ andC* are as defined for (I) in claim 1, in the form of a pure enantiomeror a mixture of isomers in any proportion, is treated with an alcohol offormula R--OH in which R is a (C₁ -C₄)alkyl, in acidic medium, in orderto form an intermediate imidate which is reacted with an amine offormula HNR₇ R₈ (III) or a diamine of formula H₂ NR₆ R₈ NH₂ (IV) inwhich R₆, R₇ and R₈ have the definitions given for (I) in claim 1: b2)the compound thus obtained, of formula: ##STR158## is coupled eitherwith a compound of formula: ##STR159## in which R₂, R₃ and R₉ are asdefined for (I) in claim 1 and Pr is a protecting group, then, afterdeprotection of the amine in acidic medium, a sulphonyl halide offormula R₁ SO₂ Hal in which R₁ is as defined for (I) in claim 1 and Halis a halogen is reacted; or with a compound of formula: ##STR160## inwhich R₁, R₂, R₃ and R₉ are as defined for (I) in claim 1: c2) thecompound of formula (1) thus obtained is isolated in base form or insalt form; d2) where appropriate, another salt of the compound offormula (I) is prepared.
 10. A pharmaceutical composition comprising, asactive principle, a compound according to claim
 1. 11. A compoundaccording to claim 6 having (R,R) isomerism on the C*-labelled carbonatoms.
 12. A compound according to claim 6 in which R_(1a) is 2-quinolyland R_(2a) is phenyl.
 13. A compound according to claim 12 having (R,R)isomerism on the C*-labelled carbon atoms.
 14. A pharmaceuticalcomposition comprising, as active principle, a compound according toclaim
 2. 15. A pharmaceutical composition comprising, as activeprinciple, a compound according to claim
 3. 16. A pharmaceuticalcomposition comprising, as active principle, a compound according toclaim
 4. 17. A pharmaceutical composition comprising, as activeprinciple, a compound according to claim
 5. 18. A pharmaceuticalcomposition comprising, as active principle, a compound according toclaim
 6. 19. A pharmaceutical composition comprising, as activeprinciple, a compound according to claim
 7. 20. A pharmaceuticalcomposition comprising, as active principle, a compound according toclaim
 11. 21. A pharmaceutical composition comprising, as activeprinciple, a compound according to claim
 12. 22. A pharmaceuticalcomposition comprising, as active principle, a compound according toclaim
 13. 23. A method for the treatment or prevention of diseasesinvolving the bradykinin receptors which comprises administering to apatient in need of such treatment an effective amount of a compoundaccording to claim
 1. 24. A method for the treatment or prevention ofdiseases involving the bradykinin receptors which comprisesadministering to a patient in need of such treatment an effective amountof a compound according to claim
 12. 25. A method for the treatment orprevention of diseases involving the bradykinin receptors whichcomprises administering to a patient in need of such treatment aneffective amount of a compound according to claim
 13. 26. A method forthe treatment or prevention of diseases involving the bradykininreceptors which comprises administering to a patient in need of suchtreatment an effective amount of a compound according to claim
 14. 27. Amethod for the treatment or prevention of diseases involving thebradykinin receptors which comprises administering to a patient in needof such treatment an effective amount of a compound according to claim5.
 28. A method for the treatment or prevention of diseases involvingthe bradykinin receptors which comprises administering to a patient inneed of such treatment an effective amount of a compound according toclaim
 6. 29. A method for the treatment or prevention of diseasesinvolving the bradykinin receptors which comprises administering to apatient in need of such treatment an effective amount of a compoundaccording to claim
 7. 30. A method for the treatment or prevention ofdiseases involving the bradykinin receptors which comprisesadministering to a patient in need of such treatment an effective amountof a compound according to claim
 11. 31. A method for the treatment orprevention of diseases involving the bradykinin receptors whichcomprises administering to a patient in need of such treatment aneffective amount of a compound according to claim
 12. 32. A method forthe treatment or prevention of diseases involving the bradykininreceptors which comprises administering to a patient in need of suchtreatment an effective amount of a compound according to claim 13.